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CONTENTS
How to Order
Sample Preparation
10
Sample Vials
and Accessories
62
Filtration
80
86
Analytical Chromatography
106
Preparative Chromatography
177
Supercritical Fluid
Chromatography (SFC)
196
198
Size-Exclusion Chromatography
GPC & GFC
267
282
Alphabetical Index
328
336
Cover Image:
Oasis HLB, a true binary mixed-mode, neutral copolymer, has a unique set of physicochemical properties that
delivers the benefits of simplicity, speed, and reproducibility in SPE methods development. Its combination
of water wettability and strong reversed-phase retentionenables high-throughput sample preparation
in 96-well plate formats and engenders confidence in analytical results.
Whats New
Page 91
Page 118
Tec hnology, this HPLC column family offers the widest range of
Waters Positive
Pressure-96 Processor
Page 53
Page 51
constant pressure, even if all the plate well positions are not filled.
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Whats New
Protein-Pak Hi Res
Ion-Exchange (IEX) Columns
Page 223
Page 217
applying the power of UPLC technology, you can now obtain accurate,
Page 196
Page 7
laboratory-scale purification.
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LITERaTURE ShOWCaSE
ACQUITY UPLC
Column Strategies Brochure
Lit. Code 720003271EN
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LITERATURE SHOWCASE
Waters DisQuE
Products for QuEChERS,
Lit. Code 720003048EN
Sample Preparation
Solutions Brochure
Lit. Code 720000848EN
Bioseparations and
Analyses Catalog
Lit. Code 720002148EN
Environmental
Analysis Catalog
Lit. Code 720002639EN
Environmental Analysis
Solutions Brochure
Lit. Code 720002766EN
Environmental Chromatography
Methods Guide
Lit. Code 720002543EN
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Waters Carbohydrate
Method Selection Tool
www.waters.com/amidemethod
www.waters.com/filterselector
LITERATURE SHOWCASE
Waters is committed to education and training. Learn from the best! Our expanding series of
easy-to-read, well-illustrated, high-quality primers, written by experts, introduce, inform,
and explain the latest technologies in analytical science.
The Quest for Ultra Performance in Liquid
Chromatography: Origins of UPLC Technology
From the dawn of LC to the present day, drawn almost entirely from original sources and first-person accounts, this
text reviews the first century of LC, showing how early the concepts of ultra performance were recognized and how
many decades it took to reduce them to practice. An extensive glossary is included.
Paperback, 54 pages,
ISBN 978-1-879732-05-6
715002098
715001531
715002099
715002531
715001940
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6
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How to order
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[ Sample Preparation ]
The Importance of Sample Preparation ..................................................................................................................................................... 12
Sorbent Selection Guide ............................................................................................................................................................................ 13
Oasis Solid-Phase Extraction (SPE) Products
Introduction ................................................................................................................................................................................. 16
Oasis MCX for Basic Compounds ............................................................................................................................................... 18
Oasis MAX for Acidic Compounds ............................................................................................................................................. 19
Oasis WCX for Strong Basic Compounds................................................................................................................................... 21
Oasis WAX for Strong Acidic Compounds ................................................................................................................................ 22
Oasis HLB for Reversed-Phase SPE ............................................................................................................................................ 23
Oasis Elution Plates.................................................................................................................................................................. 26
Oasis 96-Well High-Throughput Extraction Plates ................................................................................................................... 28
Sensitivity and Precision ........................................................................................................................................................... 30
Sorbent Amount and Solvent Selection
for the Generic Method ............................................................................................................................................................... 31
Automation ................................................................................................................................................................................. 31
Oasis On-Line Columns .............................................................................................................................................................. 32
Oasis Glass Cartridges for PPT Detection Levels ...................................................................................................................... 33
Oasis Symbiosis On-Line Cartridges ......................................................................................................................................... 33
Oasis Product Selection Guide ................................................................................................................................................... 34
Oasis Sorbent Selection Tools ................................................................................................................................................... 35
Ordering Information ................................................................................................................................................................. 36
NEW Waters Positive Pressure -96 Processor ........................................................................................................................... 38
Sep-Pak Solid-Phase Extraction (SPE) Products
Introduction ................................................................................................................................................................................. 39
Sep-Pak Sorbent Selection Guide .............................................................................................................................................. 40
Sep-Pak Cartridge Selection Guide ............................................................................................................................................ 42
Sep-Pak 96-well Plates .............................................................................................................................................................. 42
Sep-Pak 96-well Elution Plate................................................................................................................................................. 43
Sep-Pak Specialty Chemistries ................................................................................................................................................... 44
Sep-Pak DNPH-Silica Cartridges......................................................................................................................................... 45
Ozone Scrubber Cartridges ................................................................................................................................................ 46
Waters XPoSure Cartridges ................................................................................................................................................ 46
PoraPak RDX Cartridges ...................................................................................................................................................... 47
Sep-Pak Dry SPE Cartridges................................................................................................................................................ 47
Certified Sep-Pak Cartridges ...................................................................................................................................................... 48
Certified Sep-Pak Sorbent Selection Guide ............................................................................................................................... 49
Waters Extraction Manifold ........................................................................................................................................................ 50
NEW Waters Positive Pressure -96 Processor ........................................................................................................................... 51
Environmental and Food Solution Kits ..................................................................................................................................................... 52
NEW Ostro Sample Preparation Products.................................................................................................................................................. 53
DisQuE Dispersive Sample Preparation Kit ............................................................................................................................................... 54
PoraPak RXN Cartridges for Post Synthesis Cleanup................................................................................................................................ 56
10
[ Filtration ]
Sirocco Protein Precipitation Plate............................................................................................................................................................ 80
Pall Life Sciences Sample and Solvent Filtration Products ..................................................................................................................... 81
Filter Design and Membrane Choices........................................................................................................................................................ 82
Part Number Cross Reference .................................................................................................................................................................... 83
Solvent Filtration Membranes ................................................................................................................................................................... 83
Solvent Filtration Apparatus ..................................................................................................................................................................... 83
Syringe Filters Product List ....................................................................................................................................................................... 84
11
SaMPLE PREPaRaTION
lower cost
no emulsion problems
greater recoveries
fewer steps
no transporting of
samples to lab
less apparatus
faster protocol
fewer steps
reduced harm to
labile samples
greater safety
minimal evaporation
in solution
greater accuracy
no cross contamination
12
Sample Preparation
www.waters.com
SaMPLE PREPaRaTION
Air
Sorbent/Surface
Petroleum
Common
Sample Matrices
Biological
Description
Soil
Water
Food
Typical Applications
Properties
Reversed-Phase
Oasis HLB
N-VinylpyrrolidoneDVB copolymer
Sep-Pak tC18
Silica
SiC18H37
Sep-Pak C18
Silica
Si(CH3)2C18H37
Sep-Pak C8
Silica
Si(CH3)2C8H17
Sep-Pak tC2
Silica
SiC2H5
Antibiotics in cosmetics
Protein and peptide isolation by HIC
[ hydrophobic-interaction chromatography]
Reversed- or Normal-Phase
Sep-Pak
Aminopropyl
Silica
Si(CH2)3NH2
Sep-Pak
Cyanopropyl
Silica
Si(CH3)(CH2)3CN
Sep-Pak Diol
Silica
Si(CH2)3OCH2CH(OH)
CH2OH
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Sample Preparation
13
SaMPLE PREPaRaTION
Sorbent/Surface
Common
Sample Matrices
Description
Typical Applications
Properties
Normal-Phase
Sep-Pak Silica
SiO2
Sep-Pak Florisil
MgOSiO2
Sep-Pak Alumina
(A,N,B)
Al2O3
Dispersive
DisQuE
Specific Application
Oasis WCX
N-Vinylpyrrolidone
DVB copolymer
COOH
Sep-Pak Accell
Plus CM
Acrylic acid/acrylamide
copolymer on diol-silica
COO Na+
Oasis MAX
N-Vinylpyrrolidone
DVB copolymer
CH2N(CH3)2C4H9+
Sep-Pak Accell
Plus QMA
Acrylamideacrylic acid
copolymer bound
to diol-silicaC(O)
NH(CH2)3N(CH3)3+ Cl-
Oasis WAX
N-VinylpyrrolidoneDVB
copolymerCH2piperazine
Sep-Pak PSA
SiC2H4NHC2H4NH2
14
Sample Preparation
www.waters.com
SaMPLE PREPaRaTION
Sorbent/Surface
Common
Sample Matrices
Description
Typical Applications
Properties
Quantity: 1.4 g KI
Capacity: 4.2 mmoles
ozone/cartridge [theoretical]
General purpose
Quantity: 2.85 g
anhydrous Na2SO4
T heoretical capacity: 3.6 g H2O
Particle size: 80 m
Quantity: 265 mg/cartridge
Activated carbon
Particle size: 85 m
Quantity: 400 mg/cartridge
Sep-Pak Carbon
Black/Aminopropyl
Diphenylhydrazine coated
on silica
Sep-Pak X PoSure
Aldehyde Sampler
Diphenylhydrazine coated
on silica
Sep-Pak Ozone
Scrubber
Potassium iodide
Sep-Pak Dry
Anhydrous Sodium Sulfate
Sep-Pak
Porapak RDX
DVBN-Vinylpyrrolidone
copolymer
Sep-Pak PS2
StyreneDVB copolymer
Sep-Pak AC2
Carbon black
Aminopropyl silica
Sep-Pak Carbon
Black/PSA
Primary-Secondary
Amine Silica
*AOAC = Association of Official Analytical Chemists; ASTM = American Society for Testing and Materials [International]; CDFA = California Department of Agriculture; EPA = U.S. Environmental Protection Agency;
JPMHLW = Japanese Ministry of Health, Labour and Welfare; JPMOE = Japanese Ministry of the Environment; NIOSH = National Institute for Occupational Safety and Health.
Matrix Key
Air
www.waters.com
Petroleum
Biological
Soil
Water
Sample Preparation
Food
15
SaMPLE PREPaRaTION
Oasis Solid-Phase Extraction (SPE) Products
A Breakthrough in SPE
Introduction
In October 1977, Waters designed the first miniature cartridge columns
(Sep-Pak cartridges) containing silica-based adsorbents for SPE.
Demands for sample preparation led to the development of a speciallydesigned polymeric sorbent that performs optimally for reversed-phase
SPE. T he Oasis HLB copolymer with unique Hydrophilic-Lipophilic Balance
is unlike traditional SPE sorbents.
Todays goals for modern solid-phase extraction (SPE) are faster throughput,
higher recovery and reproducibility, and stronger retention and selectivity.
Now SPE can outpace high-throughput techniques such as LC/MS/MS.
Unique Water-Wettable Oasis HLB Copolymer
N-VINYLPYRROLIDONE
Analytes
properly
retained
in pore
DIVINYLBENZENE
Analytes
not
retained
Dewetted Pore
If air passes through traditional
C18 cartridge the pores dry out.
When sample is loaded, the analytes are
not retained resulting in poor recoveries.
16
www.waters.com
SaMPLE PREPaRaTION
100
100
80
80
Procainamide
Acetaminophen
Ranitidine
Propranolol
Doxepin
60
40
20
0
5
Drying Time
% Recovery
% Recovery
60
40
20
10 Min.
4
Drying Time
8 Min.
Oasis HLB 1 cc/30 mg and 1 cc/100 mg C18 cartridges were conditioned on a Waters vacuum
manifold. When the methanol reached the top of the upper frit in each cartridge, vacuum was
maintained for different times to vary the cartridge drying time. The SPE protocol was then continued.
The data shown are the average of three replicate extractions.
Multiple batches of each Oasis HLB, MCX, and MAX have been successfully
used on validated bioanalytical assays in a regulated laboratory environment.
T he variable recoveries seen with the C18 sorbents, due to the drying out
effect, are often the cause for retests, reducing laboratory productivity. In
some laboratories 10% of samples are reteststhis can be reduced using
Oasis sorbents.
Also, Oasis sorbents retain polar compounds far better than bonded silica
SPE sorbents. Note the poor recovery of the polar analyte Acetaminophen
for C18. Oasis sorbents work especially well when you need to capture
metabolites (see figure above).
% Recovery
105
95
85
Recovery for:
C18 Cartridge
H O
Literature
References
O
HO
CH
N
O
H
O
N
N
CH
3X
Theobromine
2X
N
3
Acetaminophen
H
N
H3C
Salicylic acid
3
6X
OH
HO
O
HO
Catechol
15X
0
50
100
Data shown were obtained with two 3.9 x 150 mm columns, each packed with one of the sorbents, operated under
the same conditions: mobile phase: 20 mM potassium phosphate, pH 7.0/methanol (95/5 v/v); temperature: 30 C;
flow rate: 1.0 mL/min; detection: UV @ 254 n
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17
SaMPLE PREPaRaTION
Oasis MCX for Basic Compounds
60
50
Retention (k)
Base (Unionized)
40
Cation Exchange
30
20
Reversed Phase
10
9 10 11 12 13 14
Mobile Phase pH
O
Reversed-Phase
Retention
MCX sorbent
Mixed-mode Cation eXchange
Basic Analyte
(Combined)
Retention
SO3
SO3
Reversed-phase
interaction
CH 3
N
H H
O
Sulfonic-acid-cationexchange capacity:
1.0 meq/g
Cation-Exchange
Retention
CH 3
OH
Propranolol
(basic drug)
18
www.waters.com
SaMPLE PREPaRaTION
Oasis MAX for Acidic Compounds
Oasis MAX Mixed-Mode Anion eXchange and Reversed-Phase Sorbent
for High Selectivity and Sensitivity for Acidic Compounds
T he Oasis MAX sorbent has a tightly controlled ion-exchange capacity of
0.3 meq/g ensuring reproducible SPE protocols for extraction of acidic
compounds and metabolites from biological fluids. T here are no silanol
groups to complicate the retention mode or method development. T his
novel, water-wettable, polymeric sorbent is stable from pH 0 to 14, making
method development simple and fast.
MAX sorbent
Mixed-mode Anion eXchange
CH 3
+
CH 2 N
R+
Reversed-phase
retention
Quaternary-amine-
60
Retention (k)
Acid (Unionized)
30
Anion Exchange
20
Reversed Phase
10
0
9 10 11 12 13 14
Anion-Exchange
Retention
Acidic Analyte
(Combined)
Retention
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CH 3
Suprofen
(acidic drug)
Condition:
CH3OH
Equilibrate:
H2O
Load:
Spiked acidified sample
Mobile Phase pH
Reversed-Phase
Retention
50
40
COO
anion-exchange capacity:
0.25 meq/g
C4H9
CH 3
Wash 1:
5% NH4OH in H2O
Elute 1 (Wash 2):
CH3OH
Elute 2:
2% HCOOH in CH3OH
Evaporate and Reconstitute:
20% CH3OH in H2O
19
SaMPLE PREPaRaTION
High Recovery of Acidic, Basic, and Neutral Compounds Using the Generic Oasis MAX Method
Elute 1 (basic and neutral drugs)
2
I.S.
AV
AV
0
Column:
Mobile Phase:
Detector:
Temperature:
Flow Rate:
Injection Volume:
I.S.
Sample
Sample
10 Min.
10 Min.
Results (n=6)
Peak Identification
1. Nortriptyline
2. Secobarbital
3. Ketoprofen
4. Naproxen
I.S. Butylparaben
Drugs
Nortriptyline
Secobarbital
Ketoprofen
Naproxen
g/mL
5
10
5
2.5
Type of Drug
Basic (Elute 1)
Neutral (Elute 1)
Acidic (Elute 2)
Acidic (Elute 2)
% Recovery
% RSD
94.34
88.53
91.64
103.43
0.78
1.03
2.35
3.04
Literature
References
Oasis Sample Extraction Products Brochure,
Literature Reference 720001692EN
Oasis Elution Plate Brochure,
Literature Reference 720000476EN
Topics in Solid-Phase Extraction.
Part 1. Ion Suppression in
LC/MS Analysis W hite Paper,
Literature Reference 720001237EN
Sample Prep Solutions Brochure,
Literature Reference 720000848EN
20
www.waters.com
SaMPLE PREPaRaTION
Oasis WCX for Strong Basic Compounds
High Selectivity and Sensitivity for Strong Basic Compounds
Condition:
CH3OH
Equilibrate:
H2O
WCX sorbent
Weak Cation eXchange
Load:
Spiked acidified sample
COO -
Wash 1:
5% NH4OH in H2O
Weak cation-exchange
interaction
CH3
H 3C
O
O
Reversed-phase
interaction
CH3
N
CH3
Elute 2:
2% HCOOH in CH3OH
CH3
(a)
Recovery (%)
100.0
80.0
Protriptyline (H)
60.0
Valethamate (Q)
40.0
Atenolol (P)
MRM
266.9 > 144.9
20.0
0.0
0.25
120.0
MRM
306.1 > 218.9
Recovery (%)
100.0
80.0
Protriptyline (H)
60.0
Valethamate (Q)
40.0
Atenolol (P)
20.0
0.0
Atenolol 10 ng/mL
101% Recovery
1.00
Loading volume (mL)
(b)
Instruments:
Waters 2777 Sample Manager,
and Waters 1525 binary HPLC pump,
Waters Quattro Ultima
(ESI in MRM)
0.25
MRM
264 > 191.1
Valethamate 10 ng/mL
106% Recovery
Protriptyline 10 ng/mL
102% Recovery
1.00
Loading volume (mL)
SPE recoveries for three types of bases versus volume of spiked saline solution loaded onto
(a) the Oasis WCX sorbent and (b) a commercially available silica-based WCX sorbent.
H is hydrophobic base, Q is quaternary amine, and P is polar base.
www.waters.com
LC/MS/MS chromatograms of the analytes (polar base, quaternary amine, and hydrophobic base)
extracted from rat plasma with Oasis WCX 96-well Elution plate. The SPE recoveries were
calculated by comparison with postextracted spiked plasma samples.
21
SaMPLE PREPaRaTION
Oasis WAX for Strong Acidic Compounds
High Selectivity and Sensitivity for Strong Acidic Compounds
Condition:
CH3OH
Equilibrate:
H2O
WAX sorbent
Weak Anion eXchange
+
NH
SO3
Load:
Spiked and acidified sample
+
NH 2
Wash 1:
2% HCOOH in H2O
Weak anion-exchange
interaction
Reversed-phase
interaction
Elute 2:
5% NH4OH in CH3OH
Piperazine-anion-exchange
capacity: 0.6 meq/g
100
90
% Recovery
80
70
2-Naphthalenesulfonic
acid
60
Ketoprofen
Salicylic Acid
50
40
HPLC Conditions:
Column:
Mobile Phase A:
Mobile Phase B:
Flow Rate:
Gradient:
Saline. pH = 7
Saline, pH = 1.3
Injection Volume:
Temperature:
Instrument:
100
100
2-Naphthalenesulfonic
acid
% Recovery
60
Ketoprofen
40
Salicylic Acid
20
DEA
PSA
Oasis
WAX
1 cc/
1 cc/
1 cc/ 100 mg 100 mg
30 mg
Oasis
WAX
1 cc/
30 mg
DEA
PSA
1 cc/
1 cc/
100 mg 100 mg
22
O
CH2SO3H
(1S)-Camphor-10-sulfonic acid
MW 232.3, C10H16O4S
Cone (V)
60
CID (eV)
30
108.8% recovery
%
1.29
2.75 3.10
1
100
1.59
Spiked Saline,
pH = 7
SPE recoveries for three types of acidic compounds versus volume of spiked
saline solution and rat plasma loaded onto (a) the Oasis WAX sorbent and
(b) a commercially available silica-based WAX sorbent.
CH3
Concentration: 10 pg/L
80
1.59
150 C
350 C
50 L/Hr
600 L/Hr
2.2e-3 bar (Ar gas)
CH3
MRM Transition
m/z 231.1 _ 79.8
MS/MS Conditions:
Waters Quattro Premier
ESISource Temp:
Desolvation Temp:
Cone Gas Flow:
Desolvation Gas Flow:
Collision Cell:
4.20
4.33
Concentration: 50 pg/L
114.4% recovery
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5 min
www.waters.com
SaMPLE PREPaRaTION
Oasis HLB for Reversed-Phase SPE
This generic, 1-D method (1-Dimensional only the organic strength is
changed) has proven useful for a wide variety of compound types and may
be the only protocol required, reducing method development time.
O
CH
+
CH 2 N
SO3
SO3
Condition:
1 mL CH3OH
R+
CH
Equilibrate:
1 mL H2O
Load:
1 mL spiked sample**
One Simple Procedure: Many Applications on a Universal SorbentReduced Methods Development Time
100
sed sorbents
With silica-ba
need several
(C18) you would
istries to
em
ch
t
differen
ults
ac hieve the res
shown here
Acids
Neutrals
Bases
80
n=6
RSDs < 3.5%
% Recovery
60
1-D Method
40
20
Ibu
pr
of
Na en
pr
ox
Sa
en
lic
yl
ic
Ac
Su
id
lfa
Su diaz
lfa
i
m ne
er
Ac
az
in
et
e
am
in
op
he
Th
n
eo
br
Pa omi
ra ne
za
nt
hin
Th
e
eo
ph
yl
lin
Ca e
ffe
Pr
in
oc
e
ain
am
i
Ra de
ni
tid
Ox ine
yc
od
on
Pr
e
op
ra
no
Na lol
ltr
ex
on
Sa
e
lb
ut
am
ol
Do
xe
pin
www.waters.com
23
SaMPLE PREPaRaTION
For Bases
pKa 2-10:
Use Oasis MCX
For Acids
pKa 2-8:
Use Oasis MAX
Apply Protocol 1
Apply Protocol 2
Prepare Sample
Prepare Sample
Condition/Equilibrate
Load Sample
Condition/Equilibrate
Load Sample
Wash:
2% HCOOH
Wash:
5% NH4OH
Elute 1:
100% CH3OH
Weaker
Acids
Base
50
100%
3.26
3.16
2.52
2.60
3.08
100
Protein
Precipitation
2.50
100
2.522.60
100
100
1.75
2.00
2.25
2.50
2.75
3.00
3.25
2% Phospholipids
9% Phospholipids
11% Phospholipids
12% Phospholipids
3.75
4.00 Min
Strong Acid
Acid
Strong Base
Oasis sorbent selection plate and cartridge kits enable rapid development
of SPE methods for LC/MS analysis. Having all four Oasis ion-exc hange
sorbents [MCX, MAX, WAX, and WCX] in a single plate is convenient for
scouting the best ways to accomplish efficient isolation of unknowns,
zwitterionic compounds, or mixtures of analytes with different retention/
elution properties.
100% Phospholipids
3.50
Elute 2:
2% HCOOH in CH3OH
Monitoring 5 Phospholipid
MRM Transitions
Weaker
Bases
Elute 1:
100% CH3OH
Neutrals
Elute 2:
5% NH4OH in CH3OH
Note that neutral analytes can be isolated from any of the four sorbents
in the Elute 1 step of either protocol. Choose the particular ion-exchange
sorbent that is best at removing specific matrix interferences. A good
example of this is shown below.
OH
H
H
OH
Prednisone
100%
Oasis MCX
Protocol 1 Elute 2
Oasis WAX
Protocol 1 Elute 1
SPE Recovery
50
Oasis MAX
Protocol 2 Elute 2
Oasis WCX
NH
pKa = 7.3
H H
N
H
O
N
HO
pKa = 2.6
Cephalexin
[Cephalosporin antibiotic]
A difficult amphoteric analyte
Protocol 2 Elute 1
Aliquots of prepared sample processed using Oasis 2x4 Method protocol designated for each
of 4 sorbents. Eluates from Elute 1 and Elute 2 steps analyzed by LC/MS/MS.
Clearly, Oasis MCX is the sorbent of choice.
24
www.waters.com
SaMPLE PREPaRaTION
Each plasma sample was diluted [1:1, v/v] and acidified with phosphoric acid
[4% in water]. Respective aliquots were then processed using the protocol
Oasis 2x4 Method Test on MCX: Base Isolation
Oasis MCX
and the Oasis ion-mixed-mode sorbent designated by the Oasis 2x4 Method
for the corresponding sample type. LC/MS/MS analysis was used to
determine SPE recoveries.
Oasis WAX
Protocol 1
Protocol 1
Imipramine
Decanesulfonic
acid
pKa <1
3.19
2.71
pKa ~6
MRM ES+
281.2 > 85.95
4.16e6
5 Min.
O Na
5 Min.
Oasis MAX
MRM ES+
221.1 > 79.8
2.85e3
pKa ~18
Oasis WCX
Protocol 2
Protocol 2
Ibuprofen
Valethamate
pKa ~5
2.63
2.71
O
MRM ES+
205 > 161
1.34e4
OH
Br
MRM ES+
306.2 > 163
2.10e6
O
Ibuprofen [pKa = 5.2]
www.waters.com
5 Min.
5 Min.
25
SaMPLE PREPaRaTION
Oasis Elution Plates
Waters Innovative SPE Technology
Elute in as little as 25 L
Now you can confidently perform SPE cleanup and analyte enrichment of
very small sample volumes (10-25 L) up to a maximum of 375 L. T he
Waters Oasis Elution plate combines patented plate design, proven Oasis
chemistries, and straightforward protocols that deliver high analyte recovery
and clean extracts in elution volumes as low as 25 L.
Disk Technology
Vacuum
Vacuum
26
Disk Technology
>85% recovery in 25 L
25 L
75 L
75 L
75 L
100
Elution Volume
80
% Recovery
60
40
20
0
Oasis HLB
Brand E C18
Brand E UR
Brand S C18
Elution plate
N-acetylprocainamide
betamethasone
caffeine
naproxen
amitriptyline
propranolol
www.waters.com
SaMPLE PREPaRaTION
For Bases
pKa 2-10:
Use Oasis MCX
For Acids
pKa 2-8:
Use Oasis MAX
Apply Protocol 1
Apply Protocol 2
Prepare Sample
Prepare Sample
Condition/Equilibrate
Load Sample
Condition/Equilibrate
Load Sample
Wash:
2% HCOOH
Wash:
5% NH4OH
Elute 1:
100% CH3OH
Elute 2: 5% NH4OH in
60:40 CH3CN:CH3OH
Base
Elute 1:
100% CH3OH
Neutrals
Strong Acid
Elute 2: 2% HCOOH in
60:40 CH3CN:CH3OH
Acid
Strong Base
O
N
O
O
H
Imipramine (B)
pKa = 9.4, 2 ng/mL
O F
HO
F F F F
Prednisone (N)
10 g/mL
OH
F
F
Nonafluoropentanoic Acid
(SA) pKa <0.5, 200 ng/mL
Prednisone: Neutral
Ibuprofen (A)
pKa = 5.2, 200 ng/mL
Protocol 1
Protocol 2
Oasis MAX
Oasis WCX
Pre
Oasis WAX
Pre
Oasis MCX
Valethamate (QA)
pKa >12, 2 ng/mL
OH
OH
100
80
% SPE Recovery
Each plasma sample was diluted [1:1, v:v] and acidified with phosphoric
acid [4% in water]. Respective aliquots were then processed using the
protocol and the Oasis ion-mixed-mode sorbent designated by the Oasis
2x4 Method for the corresponding sample type. LC/MS/MS analysis was
used to determine SPE recoveries. T he neutral analyte was processed on
all four sorbents used.
Br
N+
60
40
20
0
)
(sb
ate
ham
let
(n)
Va
ne
iso
dn
(n)
ic
no
nta
pe
(a)
fen
pro
(n)
Ibu
ne
iso
dn
oro
id
Ac
)
(sa
www.waters.com
flu
ne
iso
dn
na
No
Pre
(b)
ine
am
i pr
(n)
Im
ne
iso
dn
Pre
Elute 1: CH3OH
Sample Preparation
27
SaMPLE PREPaRaTION
SPE device capacity is defined as the total mass of analytes and endogenous
sample components retained by the sorbent bed under loading conditions.
Breakthrough will occur when the capacity of the sorbent bed is exceeded.
T he physicochemical properties of Oasis sorbents are designed to provide
exceptionally high loading capacity, even though each well in a Waters Oasis
Elution plate contains only 2 mg of Oasis sorbent.
SPE Recovery for 200 ng/mL Imipramine and 200 ng/mL Atenolol on Oasis MCX Elution Plate
n=8
100
100
80
80
60
60
40
40
20
20
50
100 150
350 L
Imipramine
(non-polar)
% Recovery
Atenolol (polar)
% Recovery
50
100 150
350 L
40
30
20
10
0
-10
-20
-30
-40
50
350 L
analytes, including polar and basic compounds, with RSDs less than
5% (n=96).
Waters 96-Well Plate Design
1999 R&D
100 Award
Two-Stage
Well Design
Waters award-winning plate design, with five chemistry and four sorbentmass options, provides flexible high-throughput SPE in a single device.
T he Oasis 96-well plates are designed to be used on many manifold
configurations and most robotic liquid handling systems. Oasis sorbents
unique balance of hydrophobicity and water-wettability means you will
never have to worry about poor results if individual wells of the 96-well
plate dry out. As always, you can expect Oasis SPE products to perform
reliably, delivering high and reproducible recoveries for a wide range of
28
Sample Preparation
5 mg
10 mg
30 mg
60 mg
By varying frit size and/or placement, the same plate may be filled with various quantities of sorbent per well.
Our design permits optimal recoveries, even with low sorbent weights for smaller elution volumes.
www.waters.com
SaMPLE PREPaRaTION
Extraction Protocols
Protein Precipitation
Generic Method
Inject 20 L
Inject 20 L
Inject 20 L
MRM Transition:
MS Conditions
MS Instrument:
XTerra MS C18,
2.1 x 30 mm, 3.5 m
H2O + 0.5% NH4OH
ACN + 0.5% NH4OH
0.2 mL/min
5% - 95% B 1 min.
Room temperature
Waters Alliance 2795
Ion Source:
Source Temperature:
Gas Cell:
Desolvation Temperature:
Drying Gas Flow:
Cone Gas Flow:
Quattro Triple
Quadrupole
ESI (+)
150 C
2.0 e-3 bar Argon
350 C
500 L/ hr
50 L/ hr
Cone Voltage:
Collision Energy:
Capillary Voltage:
25 volts
20
3.5 Kv
Propranolol, 1 ng/mL
Protein Precipitation
3.79
4x sensitivity
without
evaporation
Oasis HLB
4.75
5.18
4.67
4x sensitivity
without
evaporation
4.48
9x sensitivity
without
evaporation
Oasis HLB
100
100
Protein Precipitation
100
3.83
299.8 226.7
259.9 154.9
278.1 232.9
263.9 190.8
17
16
100
m/z
m/z
m/z
m/z
100
Metoclopramide (IS)
Propranolol
Amitriptyline
Nortriptyline
3.83
Oasis MCX
Intensity: 1.90
25x sensitivity
without
evaporation
x105
2
0.5
www.waters.com
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0 min
Oasis MCX
0.5
1.0
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0 min
29
SaMPLE PREPaRaTION
Sensitivity and Precision
Comparison of Oasis MCX Elution Plate and Glass Fiber and
Membrane 96-well Disk PlatesUp to 18X Increase in Sensitivity
and glass fiber plates, which require a 150 L elution volume for complete
analyte recovery. T he Oasis Elution plate enables sensitivity gains and
delivers high performance without the time consuming evaporation step.
Oasis MCX Elution Plate vs. Glass Fiber and Membrane 96-well Disk Plates
Glass Fiber SCX, 96-well Plate
250 L plasma loaded
150 L elution
% %
2.5 2.5
3.0 3.0
2.0
2.542.54
100
S/N = 11
% %
-10-10
100100
Intensity:
3.03 x104
-10
2.0 2.0
2.5 2.5
2.702.70
100100
3.0 3.0
2.0
100
S/2 = 51
% %
-10-10
2.5 2.5
3.0 3.0
2.0
2.472.47
3.0
2.70
3.5 2.0
min2.0
3.0
2.47
100
2.632.63
% %
3.5 2.0
min2.0
2.5Min.
3.5 3.5
Min.
3.0
3.0 3.0
2.0
2.452.45
100
16x Sensitivity
% %
2.63
100
100100
Intensity:
1.15 x105
2.5min
3.5 3.5
min
3.0
3.5 3.5
min
2.5min
3.0
3.5 min
2.45
Intensity:
3.11 x105
S/N = 178
3.0 3.0
2.0
2.622.62
18x Sensitivity
% %
3.5 2.0
min2.0
Intensity:
1.14 x107
S/N = 357
-10
2.5 2.5
-10-10
-10
2.0
3.0 3.0
3.5 2.0
Min.2.0
2.92
-10
2.5 2.5
-10-10
2.0
3.0 3.0
2.5 2.5
6x Sensitivity
% %
100100
Intensity:
7.14 x104
S/N = 40
3.5 2.0
min2.0
3.0
-10
2.5 2.5
-10-10
3.5 3.5
min
2.5min
2.5min
3.5 3.5
min
100100
Intensity:
7.30 x104
2.0
3.0 3.0
S/N = 17
% %
2.922.92
100
100100
-10-10
-10
2.5 2.5
-10-10
3.5 3.5
min
2.5min
-10
2.0 2.0
3.5 2.0
min2.0
2.54
100100
Terfenadinecarboxylate
0.5 ng/mL
3.0
2.92
-10-10
3.5 3.5
min
2.5min
Intensity:
7.44 x106
S/N = 82
% %
-10
2.0 2.0
2.92
2.92
100
100100
Intensity:
5.75 x106
S/N = 61
-10-10
Terfenadinealcohol
0.5 ng/mL
3.02
3.023.02
100
100100
Terfenadine
0.5 ng/mL
100
3.5 3.5
min
2.5min
3.0
3.5 min
2.62
Intensity:
5.04 x105
S/N = 927
-10
2.5 2.5
3.0 3.0
2.0
3.5 3.5
min
2.5min
3.0
3.5 min
Validation Assay of Terfenadine and Its Metabolites Using the Oasis MCX Elution Plate
30
www.waters.com
SaMPLE PREPaRaTION
Sorbent Amount and Solvent Selection for the Generic Method
The suggested amount of sorbent in a cartridge or a plate required for
your application, is given in the table below. Remember, because of the
increased capacity of the Oasis sorbents, you can use less sorbent than you
would normally need if you used a silica-based packing. The solvent used
for the elution step should be selected based on the polarity of the analyte.
The second table below gives a selection of elution solvents and each
solvent gives you different selectivity and elution strength.
Capacity and Elution Volume of Oasis
96-Well Plates and Cartridges
Sorbent
Per Device
Maximum
Mass Capacity
Typical
Sample Volumes
Elution
Volume
Elution Plate*
5 mg*
10 mg
30 mg
60 mg
60 to 400 g
0.15 to 1 mg
0.35 to 2 mg
1 to 5 mg
2 to 10 mg
10 to 375 L
10 to 100 L
50 to 200 L
100 L to 1 mL
200 L to 2 mL
25 L**
150 L
250 L
400 L
800 L
Tips for Selecting Elution Solvents for the Generic SPE Method (1-D)*
The elution solvent is selected based on polarity of analyte.
Relative
Elution Strength**
Solvent
Solvent Type
Methanol
Acetonitrile
Tetrahydrofuran
Acetone
Ethyl Acetate
proton donor
dipole-dipole
dipole-dipole
dipole-dipole
dipole-dipole
1.0
3.1
3.7
8.8
high
Methylene Chloride
dipole-dipole
high
Comments
disrupts H-bonding
medium polarity drugs
medium polarity drugs
medium polarity drugs
nonpolar drugs and
GC compatible
nonpolar drugs and
GC compatible
* When using solvents other than methanol, add 10-30% (of proton donor solvent like
methanol) to disrupt H-bonding on the Oasis HLB sorbent.
** High-Purity Solvent Guide. Burdick & Jackson Laboratories, Inc. Solvent Properties of
Common Liquids, L.R. Snyder, J. Chromatogr., 92, 223 (1974); J. Chromatogr. Sci.
16, 223 (1978)
Automation
Automation of Oasis Sample Extraction Cartridges and 96-Well Plates
Oasis SPE products are compatible with the following liquid handling and/or SPE automation systems:
Gilson ASPEC XL
www.waters.com
31
SaMPLE PREPaRaTION
Oasis On-Line Columns
On-Line SPE Columns for LC/MS/MS
T here are three HPLC Oasis on-line column configurations designed to fit
all your on-line-analysis needs.
All of these formats are available with the five Oasis patented sorbents
(HLB, MCX, MAX, WCX, and WAX) in a wide choice of particle sizes and
dimensions. T he Oasis on-line columns make it possible to analyze
a specific analyte in a sample matrix when combined with appropriate
Waters narrow-bore analytical columns (suc h as XSelect, XBridge,
SunFire, Atlantis , XTerra , or Symmetry columns).
Waste
10 Port Valve
2 Positions
Gradient
HPLC
Mass
Spectrometer
Narrow Bore
Analytical Column
10
1
8
7
4
Waste
9
8
7
4
Alliance HT
Mass
Spectrometer
Mass
Spectrometer
10
Alliance HT
Analytical
Column
32
515
System
Oasis
Waste
INJECTION
POSITION
515
System
Oasis
LOAD
POSITION
Analytical
Column
www.waters.com
SaMPLE PREPaRaTION
Oasis Glass Cartridges for PPT Detection Levels
Waters Oasis glass cartridges are available in 5 cc (200 mg) configuration
with Teflon frits for trace analysis at parts per trillion level. Eac h lot is
tested for the presence of bisphenol A and other phenols and phthalates,
assuring that endocrine disruptors in water samples can be analyzed to part
per trillion levels.
Endocrine Disruptors
PPT Recovery of Estrogens from River Water,
LC/MS, 5 ng/L Spike Level, n=4
Results
Recovery
% RSD
227
271
295
269
267
113
93
96
87
75
11
15
12
5
5
Recovery
% RSD
130
86
90
110
110
60
15
12
11
11
8
8
1. bisphenol A
2. 17b-estradiol
3. 17a-ethynylestradiol
4. estrone
5. diethylstilbestrol
1. dimethylphthalate
2. dirthylphthalate
3. n-nonylphenol
4. dibutylphthalate
5. benzylbutylphthalate
6. bis(ethylhexyl)phthalate
ISTD. o-terphenyl (internal standard)
Oasis HLB
C18
C2
20
60
40
80
100
% Recovery
Clonidine
Terconazole
www.waters.com
33
SAMPLE PREPARATION
Oasis Product Selection Guide
1 cc/
10 mg
1 cc/
30 mg
1 cc/30 mg
Flangeless
1 cc/30 mg
Gilson Adapter
3 cc/
60 mg
3 cc/60 mg
Flangeless
3 cc/60 mg
Gilson Adapter
3 cc/
540 mg
3 cc/540 mg
Flangeless
100/box
100/box
100/box
500/box
100/box
100/box
500/box
100/box
100/box
Oasis HLB 30 m
186000383
WAT094225
186001879
WAT058882
WAT094226
186001880
WAT058883
Oasis HLB 60 m
186004134
186003852
Sorbent
Oasis MCX 30 m
186004648
186000252
186001881
186001888
186000254
186001882
Oasis MCX 60 m
186000782
186000253
Oasis MAX 30 m
186004649
186000366
186001883
186000367
186001884
Oasis MAX 60 m
186000368
Oasis WCX 30 m
186004650
186002494
186002495
Oasis WCX 60 m
186002496
186002497
Oasis WAX 30 m
186004651
186002489
186002490
Oasis WAX 60 m
186002491
186002492
5 mg/
96-well
10 mg/
96-well
30 mg/
96-well
60 mg/
96-well
1/pk
1/pk
1/pk
1/pk
Oasis HLB 30 m
186000309
186000128
WAT058951
Oasis HLB 30 m
186001828BA
Oasis HLB 60 m
186000679
Oasis MCX 30 m
186001830BA
Oasis MCX 30 m
186000259
186000248
Oasis MAX 30 m
186001829
Oasis MCX 60 m
186000250
186000678
Oasis WCX 30 m
186002499
Oasis MAX 30 m
186000375
186000373
186001256
Oasis WAX 30 m
186002500
Oasis MAX 60 m
186001205
Oasis WCX 30 m
186002501
186002503
Oasis WAX 30 m
186002502
186002504
186003915
Description
34
Elution
96-well
Description
1/pk
www.waters.com
SaMPLE PREPaRaTION
6 cc/
150 mg
6 cc/
200 mg
6 cc/400 mg
Flangeless
6 cc/
500 mg
12 cc/
500 mg
20 cc/
1g
35 cc/
6g
Plus
225 mg
Vac RC
30 mg
Vac RC
60 mg
Glass Cartridge
5 cc/200 mg
30/box
30/box
100/500/box
30/box
20/box
20/box
10/box
50/box
50/box
50/box
30/box
186003365
WAT106202
186000382
186000381
186003379
186000115
186000116
186000117
186000118
186000132
186000683
186000256
186001216
186000261
186000255
186000776
186000777
186000778
186003516
186000380
186000369
186001855
186000372
186000371
186000370
186000865
186003517
186000378
186002498
186004646
186003518
186002493
186004647
186003519
Description
Oasis Elution Sorbent Selection Plate, 96-well
Oasis Sorbent Selection Plate,10 mg/96-well
Oasis Sorbent Selection Kit, 30 mg/1 cc cartridge
www.waters.com
Particle Size
Part No.
30 m
30 m
30 m
186004475
186003249
186003463
35
SAMPLE PREPARATION
Oasis HLB Sample Extraction Products
Qty.
Part No.
100/box
100/box
1000/box
100/box
186000383
WAT094225
186003908
186001879
30 m
500/box
186000988
30 m
500/box
WAT058882
30 m
30 m
100/box
100/box
WAT094226
186001880
30 m
500/box
WAT058883
30 m
60 m
60 m
60 m
30 m
60 m
60 m
60 m
60 m
60 m
60 m
30 m
30 m
30 m
60 m
30 m
30 m
30/box
100/box
100/box
100/box
30/box
30/box
30/box
20/box
20/box
10/box
50/box
50/box
50/box
50/box
30/box
96/box
96/box
48/box
12/box
10/box
10/box
10/pk
WAT106202
186003849
186004134
186003852
186003365
186003379
186000115
186000116
186000117
186000118
186000132
186005125
186000382
186000381
186000683
186001196
186003925
WAT011390
186005587
WAT054260
WAT048160
405000934
5 m
5 m
5 m
5 m
5 m
15 m
15 m
15 m
15 m
15 m
25 m
25 m
25 m
25 m
25 m
25 m
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
1/pk
5/pk
186002034
186002037
186002040
186001413
186002043
186002035
186002038
186002041
186001414
186002044
186001792
186002036
186000706
186002039
186002042
186002045
186000262
WAT046910
WAT082745
WAT084560
WAT005139
30 m
30 m
30 m
30 m
60 m
1/pk
1/pk
1/pk
1/pk
1/pk
186001828BA
186000309
186000128
WAT058951
186000679
Description
Oasis HLB cartridge
1 cc/10 mg
Oasis HLB cartridge
1 cc/30 mg
Oasis HLB cartridge
1 cc/30 mg
Oasis HLB flangeless cartridge
1 cc/30 mg
Oasis HLB cartridge with
1 cc/10 mg
Gilson ASPC adapter
Oasis HLB cartridge with
1 cc/30 mg
Gilson ASPC adapter
Oasis HLB cartridge
3 cc/60 mg
Oasis HLB flangeless cartridge
3 cc/60 mg
Oasis HLB cartridge with
3 cc/60 mg
Gilson ASPC adapter
Oasis HLB cartridge
6 cc/200 mg
Oasis HLB
3 cc/400 mg
Oasis HLB cartridge
3 cc/540 mg
Oasis HLB flangeless cartridge
3 cc/540 mg
Oasis HLB cartridge
6 cc/150 mg
Oasis HLB cartridge
6 cc/150 mg
Oasis HLB cartridge
6 cc/500 mg
Oasis HLB cartridge
12 cc/500 mg
Oasis HLB cartridge
20 cc/1 g
Oasis HLB cartridge
35 cc/6 g
Oasis HLB Plus cartridge
225 mg
Oasis HLB Light cartridge
30 mg
Oasis HLB Vac RC cartridge
20 cc/30 mg
Oasis HLB Vac RC cartridge
20 cc/60 mg
Oasis HLB glass cartridge
5 cc/200 mg
Oasis HLB Symbiosis cartridge*
1.0 x 10 mm
Oasis HLB Symbiosis cartridge*
2.0 x 10 mm
Reservoir 30 cc for Oasis cartridges
Reservoir 60 cc for Oasis cartridges
Reservoir adapter for 1 cc, 3 cc, 6 cc cartridges
Reservoir adapter for 12 cc, 20 cc, 35 cc cartridges
Reservoir adapter for 5 cc cartridges, Teflon
36
2 mg/96-well
5 mg/96-well
10 mg/96-well
30 mg/96-well
60 mg/96-well
1cc/10mg
1 cc/30 mg
1 cc/30 mg
1 cc/60 mg
3 cc/60 mg
3 cc/60 mg
3 cc/60 mg
6 cc/150 mg
6 cc/150 mg
6 cc/500 mg
20 cc/1 g
35 cc/6 g
225 mg
20 cc/60 mg
20 cc/60 mg
10 x 1 mm
Particle
Size
30 m
30 m
30 m
60 m
30 m
30 m
60 m
30 m
60 m
60 m
60 m
60 m
60 m
30 m
60 m
30 m
2.1 x 20 mm
2.1 x 20 mm
3.0 x 20 mm
3.9 x 20 mm
4.6 x 20 mm
30 m
30 m
30 m
30 m
30 m
1/pk
1/pk
1/pk
1/pk
1/pk
186002046
186002051
186002047
186002048
186002049
96-well
10 mg/96-well
30 mg/96-well
30 mg/96-well
60 mg/96-well
30 m
30 m
30 m
60 m
60 m
1/pk
1/pk
1/pk
1/pk
1/pk
186001830BA
186000259
186000248
186000250
186000678
Description
Qty.
Part No.
100/box
100/box
100/box
100/box
100/box
100/box
100/box
30/box
30/box
30/box
20/box
10/box
50/box
50/box
50/box
96/box
186004648
186000252
186001881
186000782
186000254
186001882
186000253
186000256
186000255
186000776
186000777
186000778
186003516
186000261
186000380
186002098
Literature
References
Oasis Sample Extraction Products Brochure,
Literature Reference 720001692EN
Oasis Elution Plate Brochure,
Literature Reference 720000476EN
Topics in Solid-Phase Extraction.
Part 1. Ion Suppression in
LC/MS Analysis W hite Paper,
Literature Reference 720001237EN
Sample Prep Solutions Brochure,
Literature Reference 720000848EN
Oasis WAX Sorbent for UPLC/MS
Determination of PFOS and Related
Compounds in Waters and Tissue,
Literature Reference 720001871EN
www.waters.com
SAMPLE PREPARATION
Oasis MAX Sample Extraction Products (Anion Exchange)
1 cc/10 mg
1 cc/30 mg
1 cc/30 mg
3 cc/60 mg
3 cc/60 mg
3 cc/60 mg
6 cc/150 mg
6 cc/150 mg
6 cc/500 mg
225 mg
20 cc/30 mg
20 cc/60 mg
20 cc/60 mg
10 x 1 mm
Particle
Size
30 m
30 m
30 m
30 m
60 m
30 m
30 m
60 m
60 m
60 m
30 m
30 m
60 m
30 m
2.1 x 20 mm
2.1 x 20 mm
3.0 x 20 mm
3.9 x 20 mm
4.6 x 20 mm
30 m
30 m
30 m
30 m
30 m
Description
2 mg/96-well
10 mg/96-well
30 mg/96-well
60 mg/96-well
60 mg/96-well
Qty.
Part No.
100/box
100/box
100/box
100/box
100/box
100/box
30/box
30/box
30/box
50/box
50/box
50/box
50/box
96/box
186004649
186000366
186001883
186000367
186000368
186001884
186000369
186000370
186000865
186003517
186000372
186000371
186000378
186002099
1/pk
1/pk
1/pk
1/pk
1/pk
186002052
186002057
186002053
186002054
186002055
1/pk
1/pk
1/pk
1/pk
1/pk
30 m
30 m
30 m
60 m
186001829
186000375
186000373
186001256
186001205
Part No.
96-well
30 m
186003249
96-well
30 m
186004475
1 cc/30 mg
30 m
186003463
Description
Oasis sorbent selection plate
3 rows each:
MCX, MAX, WCX and WAX
Oasis Elution Sorbent
Selection Plate, 3 rows each:
MCX, MAX, WCX and WAX
Oasis sorbent selection
cartridge kit, 10 each:
MCX, MAX, WCX and WAX
1 cc/10 mg
1 cc/30 mg
3 cc/60 mg
6 cc/150 mg
1 cc/30 mg
3 cc/60 mg
6 cc/500 mg
225 mg
2 mg/96-well
10 mg/96-well
30 mg/96-well
1.0 x 10 mm
Particle
Size
30 m
30 m
30 m
30 m
60 m
60 m
60 m
60 m
30 m
30 m
30 m
30 m
2.1 x 20 mm
3.9 x 20 mm
30 m
30 m
Description
Qty.
Part No.
100/box
100/box
100/box
30/box
100/box
100/box
30/box
50/box
1/pk
1/pk
1/pk
96/box
186004650
186002494
186002495
186002498
186002496
186002497
186004646
186003518
186002499
186002501
186002503
186002892
186002505
186002507
1 cc/10 mg
1 cc /30 mg
3 cc /60 mg
6 cc /150 mg
1 cc /30 mg
3 cc /60 mg
6 cc/500 mg
225 mg
96-well
10 mg/96-well
30 mg/96-well
60 mg
1.0 x 10 mm
Particle
Size
30 m
30 m
30 m
30 m
60 m
60 m
60 m
60 m
30 m
30 m
30 m
30 m
30 m
2.1 x 20 mm
3.9 x 20 mm
30 m
30 m
Description
Qty.
Part No.
100/box
100/box
100/box
30/box
100/box
100/box
30/box
50/box
1/pk
1/pk
1/pk
1/pk
96/box
186004651
186002489
186002490
186002493
186002491
186002492
186004647
186003519
186002500
186002502
186002504
186003915
186002893
186002508
186002509
www.waters.com
Part No.
176001835
176001836
37
SAMPLE PREPARATION
Accessories for Extraction Cartridge Manifold
Qty.
1/box
Part No.
186001831
WAT097944
WAT097945
WAT097946
Extraction Plate
Manifold
186001831
Collection Plate
30 cc Reservoir
60 cc Reservoir
Reservoir Adapters for 1, 3 and 6 cc VAC
Reservoir Adapters for 12, 20 and 35 cc VAC
Male-Male Adapter
Male Luer Plugs
Female Luer Plugs
Qty.
Part No.
48/pk
12/pk
12/pk
10/pk
100/pk
WAT011390
186005587
WAT054260
WAT048160
WAT024310
WAT044395
WAT044385
Description
Description
Qty.
Part No.
1/pk
1/pk
1/pk
1/pk
5/pk
48/pk
12/pk
100/pk
12/pk
186000262
WAT046910
WAT082745
WAT084560
WAT005139
725000417
725000418
WAT011390
186005587
WAT024310
WAT054260
10/pk
WAT048160
Qty.
Part No.
25/box
50/box
50/box
50/pk
WAT058942
WAT058943
WAT058958
WAT058959
725000417
725000418
186003522
186005234
T he Waters Positive Pressure-96 Processor offers stat of the art operation for
96-well and 1 cc flangeless cartridge formats. Each of the 96 holes in the
processor is restricted in order to maintain constant pressure, even if all the
plate well positions are not filled. Positive pressure processing offers many
advantages over traditional methods, including:
Highly reproducible
assays
Easy-to-use design
Part No.
WAT200677
WAT200606
WAT200607
WAT200608
WAT200609
Description
Waters Extraction
Manifold
38
Qty.
Part No.
1
1
1
1
186005521
186005523
186005522
405006528
186005524
186005586
www.waters.com
SaMPLE PREPaRaTION
Sep-Pak Sample Extraction Products
T he most referenced and widely used sample preparation technology
Sep-Pak bonded silica devices are recognized throughout the world and
remain the most referenced solid phase extraction product for sample
preparation. A diverse selection of formats and sorbents make Sep-Pak SPE
products ideally suited for all types of samples for GC, HPLC, and UPLC
analysis methods.
Formats:
Cartridges
96-well plate
Elution plate
Chemistries:
Reversed-phase (silica-based)
Normal-phase
Silicapolar surface used to adsorb analytes from non-polar solvents
Alumina (A, B & N) a highly active grade of alumina that is available
in acidic, basic and neutral surface chemistries
Florisilpolar, highly active, weakly basic sorbent for adsorption of
low to moderate polarity species from nonaqueous solutions
Ion-exchange (silica-based)
Specialty
PoraPak RDXfor analysis of explosives in
ground and surface water, EPA-8330
Sep-Pak Dryanhydrous Na2SO4 for removal
of residual water from non-aqueous extracts
DNPH-Silicafor air analysis of aldehydes and
ketones, EPA-TO-11A, ASTM D-5791
X PoSurefor indoor air monitoring of aldehydes and ketones
AC2activated carbon used to concentrate pesticides and herbicides
PS2styrene-divinyl benzene polymer used to concentrate
pesticides and herbicides
Carbon Black / Amino Propylfor pesticides from food
Carbon Black / PSAfor concentrating pesticides from food
www.waters.com
39
SaMPLE PREPaRaTION
Sep-Pak Sorbent Selection Guide
Reversed Phase
C18
Si(CH3)2C18H37
tC18
SiC18H37
C8
Si(CH3)2C8H17
tC2
SiC2H5
Description
Applications
Chemistry
1.
2.
3.
4.
Similar to C18
1.
2.
3.
4.
1.
2.
3.
4.
1.
2.
3.
4.
Description
Applications
Chemistry
1.
2.
3.
4.
Drugs
Drug metabolites
Pesticides
1.
2.
3.
4.
1.
2.
3.
4.
4. pH range: 2-9
1. Particle sizes:
37-55 m
5. IEX capacity:
1.75 meg/g
2. Pore size: 60
3. Surface area: 450 m2/g
Aminopropyl
Si(CH2)3NH2
Cyanopropyl
Si(CH3)(CH2)3(CN)
Diol
Si(CH2)3OCH2CH(OH)CH2OH
PSA
SiC2H4NHC2H4NH2
40
www.waters.com
SAMPLE PREPARATION
Normal Phase
Silica
SiO2
Alumina
(A, B & N)
Al2O3
Florisil
MgOSiO2
Description
Applications
Chemistry
1.
2.
3.
4.
1.
2.
3.
Description
Applications
Chemistry
1. Particle Size:
37-55 m
2. Pore Size: 300
3. Carbon: 6%
4. Counter Ion: Cl
5. pH Range 2-9
6. Loading Capacity:
200 mg
BSA/gram sorbent
7. Small Molecule
loading:
1.8-2.8 meq/gram
8. Ligand Density:
220 moles/g
1. Particle Size:
37-55 m
2. Pore Size: 300
3. Carbon: 5.5%
4. Counter Ion: Na
5. pH Range 2-9
6. Loading Capacity:
175 mg Cytochrome
C/gram sorbent
7. Small Molecule
loading:
3.1-4.2 meq/gram
8. Ligand Density:
350 moles/g
Ion Exchange
Accell Plus CM
Weak Cation-Exchanger
CO -Na+
2
Literature
References
A Sample Preparation Primer
and Guide to Solid-Phase
Extraction Methods Development,
Literature Reference WA20300
Sample Prep Solutions Brochure,
Literature Reference 720000848EN
www.waters.com
41
SaMPLE PREPaRaTION
Sep-Pak Cartridge Selection Guide
Plus
Short
Sorbent
C18
tC18
C8
tC2
Silica
Florisil
Accell Plus CM
Accell Plus QMA
Diol
Vac 1 cc/
100 mg
Vac RC/
100 mg
50/box
50/box
50/box
100/box
100/box
50/box
Part No./
Mass/Volume*
Part No./
Mass/Volume*
Part No./
Volume*
Part No./
Volume*
Part No./
Volume*
WAT020515
360 mg/0.7 mL
WAT036810
400 mg/0.8 mL
WAT036775
400 mg/0.8 mL
WAT052720
400 mg/0.8 mL
WAT023635
820 mg/1.6 mL
WAT036800
900 mg/1.4 mL
WAT023501
130 mg/0.3 mL
WAT036805
145 mg/0.4 mL
WAT036770
145 mg/0.4 mL
WAT052725
145 mg/0.4 mL
WAT023537
120 mg/0.4 mL
WAT023543
145 mg/0.3 mL
WAT023531
130 mg/0.4 mL
WAT023525
130 mg/0.4 mL
WAT023549
280 mg/0.35 mL
WAT023555
280 mg/0.35 mL
WAT023561
280 mg/0.35 mL
WAT023513
130 mg/0.3 mL
186004578
130 mg/0.3 mL
WAT023507
130 mg/0.3 mL
WAT023519
130 mg/0.4 mL
WAT051910
360 mg/0.85 mL
WAT054955
0.13 mL
WAT054960
0.11 mL
WAT054965
0.11 mL
WAT023590
0.2 mL
WAT036820
0.25 mL
WAT036785
0.25 mL
WAT052710
0.25 mL
WAT023595
0.25 mL
WAT023600
0.2 mL
WAT023625
0.25 mL
WAT023620
0.25 mL
WAT023575
0.1 mL
WAT023580
0.1 mL
WAT023585
0.1 mL
WAT023610
0.2 mL
186004561
0.2 mL
WAT023615
0.2 mL
WAT023605
0.25 mL
WAT036935
0.2 mL
WAT043410
0.25 mL
WAT043415
0.25 mL
WAT020520
690 mg/1.6 mL
WAT020525
910 mg/1.4 mL
WAT020550
360 mg/0.8 mL
WAT020545
360 mg/0.8 mL
Vac 1 cc/
50 mg
Part No./
Mass/Volume*
Alumina B
PSA
Classic
Short/Long
50/box
Plus
Light
Part No./
Mass/Volume*
Alumina A
Alumina N
Plus
Long
WAT020500
1710 mg/1.2 mL
WAT020505
1710 mg/1.2 mL
WAT020510
1710 mg/1.2 mL
WAT020535
360 mg/0.7 mL
186004538
360 mg/0.7 mL
WAT020540
360 mg/0.7 mL
WAT020530
360 mg/0.8 mL
WAT051900
690 mg/2.0 mL
WAT051960
900 mg/1.7 mL
WAT010910
360 mg/1.1 mL
WAT010835
360 mg/1.1 mL
WAT051800
1850 mg/1.8 mL
WAT051820
1850 mg/1.8 mL
WAT051810
1850 mg/1.8 mL
WAT010830
360 mg/0.85 mL
186004560
360 mg/0.85 mL
WAT010823
360 mg/0.85 mL
WAT054980
0.15 mL
WAT054985
0.12 mL
186004562
0.1 mL
WAT054975
0.13 mL
WAT036940
0.25 mL
WAT043460
0.25 mL
WAT043475
0.2 mL
186004567
0.2 mL
WAT043480
0.25 mL
Part No.
42
186002319
186002320
186002321
186001917
186003966
www.waters.com
SAMPLE PREPARATION
Vac 3 cc/
200 mg
Vac 3 cc/
500 mg
Vac RC/
500 mg
Vac 6 cc/
500 mg
Vac 6 cc/
1g
Vac 12 cc/
2g
Vac 20 cc/
5g
Vac 35 cc/
10 g
50/box
50/box
50/box
30/box
30/box
20/box
20/box
20/box
Part No./
Volume*
Part No./
Volume*
Part No./
Volume*
Part No./
Volume*
Part No./
Volume*
Part No./
Volume*
Part No./
Volume*
Part No./
Volume*
WAT054945
0.42 mL
WAT054925
0.34 mL
WAT054940
0.34 mL
WAT020805
0.8 mL
WAT036815
1.0 mL
WAT036780
1.0 mL
WAT052715
1.0 mL
WAT020810
1.2 mL
WAT020815
0.8 mL
WAT020855
1.1 mL
WAT020850
1.1 mL
WAT020820
0.4 mL
WAT020825
0.4 mL
WAT020830
0.4 mL
WAT020840
0.8 mL
186004536
0.8 mL
WAT020835
0.8 mL
WAT020845
1.0 mL
WAT036945
0.8 mL
WAT043425
1.0 mL
WAT043430
1.0 mL
WAT043395
1.2 mL
WAT036790
1.1 mL
WAT054525
1.1 mL
WAT036905
2.0 mL
WAT036795
1.9 mL
WAT054570
1.9 mL
WAT052705
1.9 mL
WAT036910
1.9 mL
WAT043390
2.0 mL
WAT054590
1.9 mL
WAT054595
1.9 mL
WAT054580
0.8 mL
WAT054585
0.8 mL
WAT054575
0.8 mL
WAT054605
2.0 mL
186004537
2.0 mL
WAT054600
2.0 mL
WAT054610
1.9 mL
WAT036915
3.6 mL
WAT043380
3.5 mL
WAT054615
3.5 mL
WAT036925
8.0 mL
WAT043365
7.8 mL
WAT054660
7.8 mL
WAT043345
16.8 mL
WAT043350
16.3 mL
WAT054700
16.3 mL
WAT054930
0.53 mL
WAT054935
0.42 mL
WAT036950
1.2 mL
WAT043435
0.8 mL
WAT054505
1.1 mL
WAT054500
1.1 mL
WAT043400
1.2 mL
WAT043405
1.2 mL
WAT054545
1.2 mL
WAT054550
1.2 mL
WAT054535
0.5 mL
WAT054540
0.5 mL
WAT054530
0.5 mL
WAT054560
1.2 mL
186004563
1.2 mL
WAT054555
1.2 mL
WAT054565
1.1 mL
WAT043485
0.4 mL
WAT054515
0.8 mL
186004568
0.8 mL
WAT054520
1.0 mL
WAT036920
3.9 mL
WAT043385
3.6 mL
WAT054635
3.5 mL
WAT054640
3.5 mL
WAT054620
1.8 mL
WAT054625
1.8 mL
WAT054630
1.8 mL
WAT054650
3.6 mL
186004564
3.6 mL
WAT054645
3.6 mL
WAT054655
3.5 mL
WAT036930
11.0 mL
WAT043370
8.0 mL
WAT054675
7.8 mL
WAT054680
7.8 mL
WAT054670
3.9 mL
WAT054665
3.9 mL
WAT043375
3.9 mL
WAT054695
8.0 mL
186004565
8.0 mL
WAT054685
8.0 mL
WAT054690
7.8 mL
WAT043355
23.4 mL
WAT043360
16.8 mL
WAT054720
16.3 mL
WAT054725
16.3 mL
WAT054710
8.2 mL
WAT054715
8.2 mL
WAT054705
8.2 mL
WAT054740
16.8 mL
186004566
16.8 mL
WAT054730
16.8 mL
WAT054735
16.3 mL
*Hold up volume
www.waters.com
Part No.
186002318
43
SAMPLE PREPARATION
Sep-Pak Specialty Chemistries
Air Testing
Description
Part Number
Mass/Volume/Type
Chemistry
WAT037500
20/box
WAT039550
20/box
XPoSure
WAT047205
20/box
Ozone Scrubber
WAT054420
20/box
Description
Part Number
Mass/Volume/Type
Chemistry
WAT047220
30/box
WAT054265
50/box
1. 2.85 g anhydrous
sodium sulfate
JJAN20229
50/box
1. Particle Size: 85 m
JJAN20131
50/box
186003369
30/box
DNPH Short
DNPH Long
Potassium/Iodine
Porapak RDX
DVB Vinylpryrrolidone
Sep-Pak Dry
Sodium Sulfate
AC2
Fine Activated
Carbon
PS2
Styrene - DVB
Carbon Black/
Amino Propyl
Carbon Black/
PSA
186004590
30/box
2. 400 mg/cartridge
activated carbon
1. Particle Size: 80 m
2. 300 mg cartridge
Amine Silica
Accell Plus
QMA Carbonate
186004051
50/box
44
www.waters.com
SaMPLE PREPaRaTION
Increased safety.
www.waters.com
Qty
Part No.
20/box
20/box
WAT037500
WAT039550
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
DNPH
Formaldehyde-DNPH
Acetaldehyde-DNPH
Acetone-DNPH
Acrolein-DNPH
Propionaldehyde-DNPH
Crotonaldehyde-DNPH
Butanone-DNPH
Butyraldehyde-DNPH
Benzaldehyde-DNPH
Isovaleraldehyde-DNPH
Valeraldehyde-DNPH
o-Tolualdehyde-DNPH
m-Tolualdehyde-DNPH
p-Tolualdehyde-DNPH
Hexaldehyde-DNPH
2,5-Dimethylbenzaldehyde-DNPH
4.8 ppbv
1.2 ppbv
118.0 ppbv
0.8 ppbv
0.7 ppbv
The sample was obtained in a chemical research laboratory using a portable sampling pump.
One hundred liters of air was drawn through the cartridge at 0.65 L/min. The sample was found to
contain low concentration of formaldehyde (4.8 ppbv), acetaldehyde (1.2 ppbv) and methyl ethyl ketone
(0.8 ppbv), but a significant amount of acetone (118 ppbv). Significant concentrations of acetone are
frequently found in laboratories because of its widespread use as a solvent.
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
DNPH
Formaldehyde-DNPH
Acetaldehyde-DNPH
Acetone-DNPH
Acrolein-DNPH
Propionaldehyde-DNPH
Crotonaldehyde-DNPH
2-Butanone-DNPH
Butyraldehyde-DNPH
Benzaldehyde-DNPH
o-Tolualdehyde-DNPH
m-p-Tolualdehyde-DNPH
2,5-Dimethylbenzaldehyde-DNPH
2290 ppbv
367 ppbv
26 ppbv
8 ppbv
12 ppbv
36 ppbv
24 ppbv
15 ppbv
32 ppbv
5 ppbv
11 ppbv
3 ppbv
45
SaMPLE PREPaRaTION
Air
Sample
In
O3 Scrubber
Cartridge
DNPH-Silica
or X PoSure
Cartridge
Vacuum
Sampling
Pump
Ozone Scrubber
Description
Ozone Scrubber
Qty.
Part No.
20/box
WAT054420
Waters X PoSure Aldehyde Sampler Cartridges for Monitoring Aldehydes in Indoor Air
Based on an extension of our DNPH coating
technology, XPoSure aldehyde sampler cartridges
are t he most sensitive active samplers
available today.
1.
2.
3.
4.
5.
6.
Highest Sensitivity
Compared to existing sampling tube tec hnology whic h have high and
variable backgrounds, X PoSure cartridges are guaranteed to give
consistent low aldehyde backgrounds, cartridge-to-cartridge, lot-to-lot.
High-Collection Efficiencies
You can ac hieve > 95% collection efficiencies for all aldehydes at flows
of up to a liter per minute. And, you only need to use one cartridge
no breakthrough bed is necessary.
Low Pressure DropUse with Portable Personal Sampling Pumps
Large particle size and higher porosity frits make the X PoSure cartridge
compatible with personal sampling pumps.
DNPH
Formaldehyde-DNPH
Acetalhyde-DNPH
Acetone-DNPH
Butanone-DNPH
Isovaleraldehyde-DNPH
6
Lab Air Sample
Cartridge Blank
0
10
12
14
Min.
The above sample was collected in a chemical research laboratory using a portable sampling pump. One hundred
liters of air was drawn through the cartridge at 0.65 L/min. The chromatogram shows ormaldehyde (4.8 ppbv),
acetaldehyde (1.2 ppbv), acetone (118 ppbv), butanone (0.8 ppbv), and isovaleraldehyde (0.7 ppbv).
Easy-to-Use
Sample, elute and shoot, its that easy. Youll never have to break open and
manipulate a glass tube again. And because the cartridges are made from
high density polyethylene (HDPE), breakage is not a concern.
T he figure on t he right s hows two traces. An actual cartridge blank
demonstrating extremely low background levels and as an actual
laboratory air sample.
46
Qty.
Part No.
20/box
WAT047205
www.waters.com
SaMPLE PREPaRaTION
High Sensitivity
Unmatched Recoveries
T he specially prepared resin is highly selective for nitroaromatic and
nitramine compounds, resulting in recoveries of 90% or greater. Recovery
data from preconcentrating 500 mL of explosives standards in sterile
water at two concentrations on PoraPak Rdx Sep-Pak Vac columns. Number
of replicates = 7.
1 ppb
% Recovery
% RSD
10 ppb
% Recovery
% RSD
HMX
TNB
RDX
100.5
95.9
90.9
6.7
3.5
6.4
100.5
99.3
98.7
3.9
3.3
3.2
DNB
TNT
TETRYL
99.5
97.0
89.0
3.2
3.0
6.4
99.2
102.
102.8
3.2
3.7
4.7
NB
3,5-DNA
2,4-DNT
96.5
91.2
97.3
2.5
3.3
3.4
97.9
98.2
99.9
2.8
3.6
3.4
2,6-DNT
2-Am-DNT
4-Am-DNT
94.5
92.4
90.0
3.4
5.2
4.9
98.7
98.0
97.2
3.4
3.7
4.1
4-NT
2-NT
89.5
96.8
4.3
6.6
100.4
93.4
3.7
3.0
Compound
Explosives
Blank
Min.
Achieve the lowest method detection limits using Waters PoraPak Rdx cartridges.
The chromatogram above shows the analysis of reagent H2O spiked at the 1 ppb level.
The bottom chromatogram is a reagent (H2O) blank.
Qty.
Part No.
30/box
WAT047220
WAT023344
WAT054260
186005587
WAT024310
12/box
12/box
www.waters.com
Qty.
Part No.
50/box
WAT054265
47
SAMPLE PREPARATION
Certified Sep-Pak Solid-Phase Extraction Cartridges
Manufacturing
Our world-class manufacturing facilities strive to improve quality
expectations for SPE product performance. We manufacture under the
highest quality standard in the industry including ISO 9001:2000,
ISO 13485:2003 and current Good Manufacturing Practices (CGMP). Each
Certified Sep-Pak Product is thoroughly QC tested using:
Sorbent Specifications based on:
Contaminants including hydrocarbons and other
environmental contaminants
Sorbent functionality including:
ligand density
particle size distribution
surface activity
Chromatographic performance
Assembly Specifications based on:
Frits and barrels dimensional tolerance
Chromatographic testing of total residual extractables including:
Hydrocarbons
Plasticizers
Anti-oxidants
SPE Procedure
Scan EI+
TIC
2.00e8
TCP
1
2 3
5
9 10
15,16
13
12
11
14
17
Contaminants found in
2
UT BHT
BHA
UFA
Competitor Cartridge
Scan EI+
TIC
2.00e8
UFE
UFE
UFE
aldehyde
U. unknown
H. hydrocarbon (alkane)
Scan EI+
TIC
2.00e8
GC Analysis
Instruments:
3
8
10
11
1. -HCH (hexachlorocyclohexane)
2. -HCH
3. -HCH (lindane)
4. -HCH
5. heptachlor
12
13
6 aldrin
7. heptachlor epoxide
8. endosulfan I
9. DDE
10. dieldrin
14
15
16
17
11. endrin
12. endosulfan II
13. DDD
14. endrin aldehyde
15. endosulfan sulfate
18
19
Elute:
5 mL 90:10 hexane/acetone (v/v)
BHB. butylatedhydroxybenz-
Load:
1 mL hexane extract sample
using gravity flow
20
16. DDT
17. methoxychlor
21 min
Column:
Injection:
Temperature Program:
Evaporate:
To just under 1 mL and adjust
to 1.0 mL final volume
TCP (trichlorophenol) is added to the target compound mix as a polar probe to test sorbent specificity. Quantitative removal
of TCP is expected using Florisil sorbent under these SPE conditions.
48
www.waters.com
SaMPLE PREPaRaTION
Certified Sep-Pak Sorbent Selection Guide
C18
Silica
Alumina
(A, B, N)
A polar sorbent for analyte isolation from nonpolar solvents like hydrocarbons and less
polar esters and ethers
Florisil
Ordering Information
Description
Part No.
C18
Certified Sep-Pak Vac, 3 cc, 200 mg, 50/box
Certified Sep-Pak Vac, 3 cc, 500 mg, 50/box
Certified Sep-Pak Vac, 6 cc, 500 mg, 30/box
Certified Sep-Pak Vac, 6 cc, 1 g, 30/box
186004618
186004619
186004620
186004621
Silica
Certified Sep-Pak Vac, 3 cc, 200 mg, 50/box
Certified Sep-Pak Vac, 3 cc, 500 mg, 50/box
Certified Sep-Pak Vac, 6 cc, 500 mg, 30/box
Certified Sep-Pak Vac, 6 cc, 1 g, 30/box
186004614
186004615
186004616
186004617
Alumina A
Certified Sep-Pak Vac, 3 cc, 500 mg, 50/box
Certified Sep-Pak Vac, 6 cc, 500 mg, 30/box
Certified Sep-Pak Vac, 6 cc, 1 g, 30/box
186004602
186004603
186004604
Alumina B
Certified Sep-Pak Vac, 3 cc, 500 mg, 50/box
Certified Sep-Pak Vac, 6 cc, 500 mg, 30/box
Certified Sep-Pak Vac, 6 cc, 1 g, 30/box
186004605
186004606
186004607
Alumina N
Certified Sep-Pak Vac, 3 cc, 500 mg, 50/box
Certified Sep-Pak Vac, 6 cc, 500 mg, 30/box
Certified Sep-Pak Vac, 6 cc, 1 g, 30/box
186004608
186004609
186004610
Florisil
Certified Sep-Pak Vac, 3 cc, 500 mg, 50/box
Certified Sep-Pak Vac, 6 cc, 500 mg, 30/box
Certified Sep-Pak Vac, 6 cc, 1 g, 30/box
186004611
186004612
186004613
Literature
References
www.waters.com
Environmental Chromatography
Methods Guide,
Literature Reference 720002543EN
Food Safety
Application Notebook,
Literature Reference 720002565EN
49
SAMPLE PREPARATION
Waters Extraction Manifold for Use With Solid-Phase Extraction Cartridges
T he vacuum manifold has the capacity to process up to twenty samples
simultaneously. T he extraction manifold has enhanced features designed
for use with conventional silica-based, solid-phase extraction cartridges
as well as modifications that allow you take full advantage of the unique
performance characteristics of our Oasis HLB extraction cartridges.
Just look at what this manifold has to offer:
A Precision-machined Delrin cover with alignment posts
for quick and easy alignment with test tube rack.
B Vacuum gauge placement on cover, not in fluid path allows
for quick and easy waste removal at bottom by vacuum.
C
Description
Part No.
WAT200677
WAT200606
WAT200607
WAT200608
WAT200609
Qty.
Part No.
WAT200685
WAT200691
WAT200686
WAT200687
WAT200688
WAT200689
WAT200690
WAT058839
WAT058851
WAT058840
WAT058871
186005234
WAT200678
WAT200679
WAT200680
WAT200681
WAT011390
186005587
WAT024310
WAT054260
WAT048160
WAT085114
WAT085115
50
www.waters.com
SAMPLE PREPARATION
Part No.
48/pk
12/pk
12/pk
10/pk
100/pk
WAT011390
186005234
WAT054260
WAT048160
WAT024310
WAT044395
WAT044385
Description
Part No.
WAT011400
Description
Waters Positive Pressure-96 Processor
96-flangeless cartridge holder
96-place sealing gasket
Elution positive pressure spacer
Gas supply adapter, includes 1/8 in to 1/4 in NPT
fitting, 6 feet of 1/4 in tubing
10 mL x 24 waste collection plate
www.waters.com
Qty.
Part No.
1
1
1
1
186005521
186005523
186005522
405006528
186005524
186005586
Easy-to-use design
Literature
References
Waters Positive Pressure-96
Processor Brochure,
Literature Reference 720003453EN
Bioanalysis Brochure,
Literature Reference 720003692EN
51
SAMPLE PREPARATION
Environmental and Food Solution Kits
ACQUITY PFC Column Kit
Waters Carbamate Analysis Kit for Food Testing includes a Waters Carbamate
column, Oasis HLB cartridges, vials and reference standards. Optimized for
use with AOAC methods for Carbamate Analysis, these products simplify
your analysis while increasing your confidence in the results.
Description
Part No.
176001692
Description
Carbamate Analysis Kit for Food Testing
Part No.
186004719
Waters ACQUIT Y PFC Analysis Kit includes Oasis SPE cartridges, PFC
calibration and reference standards, Certified vials, ACQUITY columns and
the necessary instrument components to optimize your instrument for trace
level detection of PFCs.
Waters EPA Method 1694 Analysis Kit includes the XTerra MS C 18,
Atlantis HILIC columns, and Oasis HLB cartridges; all of whic h are
specified in the EPA Method.
Description
EPA Method 1694 Analysis Kit
Description
Part No.
176001744
Description
Part No.
52
176001740
Part No.
176001634
Description
ACQUITY Bisphenol A Column Kit
ACQUITY Bisphenol A Method Kit
Part No.
176001955
186004932
www.waters.com
SaMPLE PREPaRaTION
NEW Ostro Sample Preparation
T he Faster Way to Cleaner
Ostro sample preparation products provide a novel solution for t he
cleanup of phospholipids in plasma and serum. Requiring minimal to
no method development and using a simple protocol, this tec hnology
can be quickly implemented in order to optimize your laboratorys
workflow. Providing cleaner, more reproducible extracts than competitive phospholipid removal devices or tec hniques, t he Ostro 96-well
plate allows for more sensitive analyses, increased sample throughput,
and reduced instrument downtime.
Protocol
Minimizing method development time, the standard Ostro protocol will provide
excellent results for a wide variety of acidic, basic and neutral compounds.
Phospholipid Removal
350000
300000
Pipette 50-200 L of
plasma into wells.
250000
200000
150000
100000
50000
0
Ostro
LLE
SSLE
Reproducibility
Comparative % removal of total phospholipids from 6 different lots
of plasma using Ostro (0.19% RSD), phospholipid removal plate from
competitor A (24.5% RSD) and phospholipid removal plate from
competitor B (40.9% RSD).
* For sample volumes 50 L or less, a higher solvent to plasma ratio may be neccessary.
Qty.
Part No.
186005518
100
90
% Phospholipid Removal
80
70
60
50
40
Literature
References
30
20
10
0
Ostro
www.waters.com
Competitor A
Competitor B
Bioanalysis Brochure,
Literature Reference 720003692EN
53
SaMPLE PREPaRaTION
DisQuE Dispersive Sample Preparation Solutions
Dispersive sample preparation, commonly referred to as QuEChERS, is a
simple and straightforward sample preparation technique suitable for multiresidue pesticide analysis in a wide variety of food and agricultural products.
Waters DisQuE Dispersive Sample Preparation Kit contains conveniently
packaged centrifuge tubes with pre-weighted sorbents and buffers designed
for use with regulatory methods.
Waters DisQuE dispersive sample preparation is a well-proven, high throughput
sample preparation method that offers several advantages over traditional
SPE techniques:
125
Baby Food
Mango
Avocado
Eliminates outsourcing
Recovery %
100
80
60
40
20
0
Pesticide
UPLC Conditions
LC System:
Column:
Column Temp:
Sample Temp:
Flow Rate:
Mobile Phase A:
Mobile Phase B:
Gradient:
MS Conditions
MS System:
Ionization Mode:
Capillary Voltage:
Desolvation Gas:
Cone Gas:
Source Temp:
Acquisition:
Collision Gas:
0.5
1.5
2.5
3.5
4.5
5.5
6.5
7.5
8 Min.
54
www.waters.com
SaMPLE PREPaRaTION
In some instances, when the sample contains excess fat or high amounts of
c hlorophyll, it is beneficial to add graphitized carbon black (GCB) or
additional C18 sorbent. T he additional sorbents are available in bulk and can
be provided for this purpose.
DisQuE Dispersive Sample Preparation Products
Description
Part No.
DisQuE Kits
Extraction Tubes
176001676
176001903
50 mL tube containing 1 g
Trisodium Citrate Dihydrate,
0.5 g Disodium Hydrogencitrate
Sesquihydrate, 1g NaCl, and
4 g MgSO4 (white cap)
186004571
186004837
Clean-Up Tubes
2 mL tube
2 mL tube
2 mL tube
containing
containing
containing
150 mg MgSO4 150 mg MgSO4, 150 mg MgSO4
and 50 mg PSA
50 mg PSA,
and 25 mg PSA
(white cap)
and 50 mg C18
(yellow cap)
(blue cap)
2 mL tube
containing
150 mg MgSO4,
25 mg PSA
and 25 mg C18
(green cap)
186004572
186004830
186004831
186004832
186004833
186004834
Bulk Sorbents
Graphitized Carbon Black. 25 g Bottle
C18, 100g Bottle
186004835
WAT035672
Literature
References
15 mL tube containing
900 mg MgSO4 and
150 mg PSA (white cap)
www.waters.com
15 mL tube containing
900 mg MgSO4, 150 mg PSA,
and 150 mg C18 (blue cap)
Determination and
Confirmation of Priority
Pesticide Residues
in Baby Food,
Literature Reference
720002812EN
55
SaMPLE PREPaRaTION
PoraPak Rxn Cartridges
for Post Synthesis Cleanup
PoraPak
TM
56
T he polymer used in PoraPak Rxn products is resistant to shrinking or swelling in the organic solvents typically used in synthetic reactions. Tests
with the following solvents demonstrate that the packed bed maintains
good flow properties:
DCE
DMF
DCM
THF
DMSO
Acetone
www.waters.com
SaMPLE PREPaRaTION
Condition
Load
Wash
Elute
Cleanup Challenges
Challenge
Solution
Load reaction mix on PoraPak Rxn CX cartridge and pass through high
boiling point solvent such as DMSO.
Wash with methanol.
Elute using 5% ammoniated methanol. Achieve sample enrichment and save time
by evaporative removal of a low boiling point solvent before the next step.
www.waters.com
57
SaMPLE PREPaRaTION
Reaction Products
Reaction Components
Technique
Recommended Product
Bases
PoraPak Rxn CX
Acids
Pass T hrough
PoraPak Rxn CX
Neutrals
Adsorption
PoraPak Rxn RP
TFA removal
PoraPak Rxn CX
PoraPak Rxn CX
Fractionate sample
before Prep for bases
PoraPak Rxn CX
PoraPak Rxn CX
Adsorption
In the catch and elute technique, the reaction mixture is loaded onto
the cartridge; the analytes of interest are retained by the sorbent. A wash
step follows to remove additional reaction components from the cartridge.
A strong solvent is used to elute the analytes from the cartridge. Sample
concentration results when the final elution volume is smaller than the
load volume.
Pass T hrough
In the pass-through technique, the sorbent does not retain the compounds
of interest, but does retain some class(es) of compounds and impurities,
that can be discarded with the column. Fractionation is achieved by the
interaction between the impurities in the reaction mixture and the sorbent.
No sample enrichment occurs during the solid-phase extraction (SPE) step.
58
www.waters.com
SaMPLE PREPaRaTION
Cleanup Procedures
Using PoraPak Rxn CX: Strong Cation-Ion Exchange Catch-and-Elute and Pass-T hrough Protocols
Catch and Elute
Pass T hrough
Condition Step
100% methanol
Condition Step
100% methanol
Load Step
Fraction collected
contains acids,
neutrals and
reaction solvent
Load Step
Fraction collected
contains acids,
neutrals and
reaction solvent
Wash Step
100% methanol
fraction contains
more impurities,
acids and neutrals
Elution Step
5% ammoniated
methanolfraction
contains bases
Wash Step
100% methanol
fraction contains
acids and neutrals
Catch-and-Elute Strategy
elute bases using 5% ammoniated methanol.
Pass-T hrough Strategy the retained compounds are unwanted
reaction components; no need for further elution step.
Recombine Load
and Wash Steps
Condition Step
Load Step
Wash Step
Generic protocol
Wash 1 Water. Fraction contains polar compounds.
Wash 2 Water:methanol (1:1). Fraction collected contains
less polar compounds.
Elution Step
www.waters.com
59
SaMPLE PREPaRaTION
3.0
2.8
2.6
2.4
2.2
2.0
1-Naphthaldehyde
C
R
H
+
R
R'
H2O
4-Methoxybenzylamine (MBA)
R'
+H2
1.8
PRODUCT
(>80% yield)
1.6
N
R
R'
H C
R
1-Naphthaldehyde
1.4
1.2
1.0
8.0e-1
N-(Naphthylmethyl)methoxybenzylamine (product)
MBA
6.0e-1
4.0e-1
2.0e-1
0.0
UPLC Conditions
Instrument:
Column:
Column Part Number:
Mobile Phase A:
Mobile Phase B:
Gradient and Flow Rate:
Injection Volume:
Column Temperature:
Detector:
UV Wavelength:
Load Step
Catch-and-Elute
Procedure
Condition Step
100% methanol
1.5
1.0
Curve
5.0e-1
Load Step
Initial
6
0.0
Wash Step
Wash Step
100% methanol
2.5
2.0
1.5
1.0
Elution Step
5% ammoniated
methanol
5.0e-1
0.0
Elution Step
The cartridge is washed with ammoniated methanol, eluting
all the bases. This fraction contains the product.
2.5
2.0
Using the catch-and-elute procedure to clean up this synthesis mixture enables a quick collection of
a limited number of fractions, increasing the certainty of finding the product rapidly. Assay the product
fraction for confirmation, then begin the evaporation of the more volatile elution solvent.
78% Product
in Eluate
1.5
1.0
5.0e-1
0.0
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
7.0 Min
Removal of TFA from Reversed Phase Prep-LC Fractions; Isolation of the Purified Free-Base Product
Removal of T FA from Prep-LC
product fraction is important
as residual T FA has been linked
to compound degradation. T his
simple protocol removes T FA
and water from product fraction
for rapid evaporation. After
evaporation the compound is in
free-base form.
Product (NMBA)
TFA
2 .5 2
Elute
2 .6 1
Wash 2
60
Elute
Wash 2
Wash 1
LC-MS(MS) chromatograms
showing product only in the
elute fraction and all TFA in
the load and wash fractions
2. 93
Wash 1
2. 92
Load
2.0
2.2
2.4
2.6
2.8
3.0 Min.
Load
1
5 Min.
www.waters.com
SAMPLE PREPARATION
Product Guidelines
T he following tables should be used as guidelines for loading capacity,
flow rates, and volumes of wash and elution solvents. T hese tables take into
consideration the dimension of the device and the bed volumes suggest
appropriate flow rates and volumes for the condition, wash and elute steps.
Maximum Capacity
Guidelines
Configuration
PoraPak Rxn RP
PoraPak Rxn CX
Retained Compounds
Retained Base
T he reaction mix can be poured into the cartridge and allowed to flow by
gravity. If the flow is too slow, simply add a small amount of pressure or
vacuum to increase it. If there is precipitate in the reaction vessel, rinse
the vessel with more loading solvent and add to the top of the cartridge.
During the washing steps, product trapped by the precipitate may be
dissolved and can be retained on the packed bed for later elution under the
right solvent conditions.
6 cc/150 mg
15 mg/50 mole
15 mg/50 mole
6 cc/400 mg
40 mg/150 mole
40 mg/150 mole
Device
20 cc/2 g
60 cc/5 g
T he capacity is for the total of all compounds in the reaction that will be
retained. For cation exchange, the capacity is for all bases in the reaction
mixture including product and reactants.
Consider smaller loads than maximum capacity guidelines or choose a
larger device to avoid compound break through.
Condition
Wash
Elute
6 cc/150 mg
3 mL
4 mL
4 mL
6 cc/400 mg
5 mL
10 mL
10 mL
20 cc/2 g
20 mL
20 mL
20 mL
60 cc/5 g
45 mL
45 mL
45 mL
T his table provides guidelines for volumes used in the condition, wash and
elution steps for the various sizes of PoraPak Rxn cartridges.
2 mL/min
20 cc cartridge
5 mL/min
60 cc cartridge
9 mL/min
Flow rate guideline is meant to provide a flow for reaction loading step.
www.waters.com
Part No.
PoraPak Rxn CX
Part No.
PoraPak Rxn RP
186004541
186004542
186004543
186004544
186004569
186004545
186004546
186004547
186004548
186004570
61
Needle design
Volatility
Chemical compatibility
Cleanliness
Sample volume
PTFE/Silicone
62
Pre-slit PTFE/Silicone
PE Septumless
Literature
References
Sample Vials & Accessories Brochure,
Literature Reference 720001818EN
Waters LC/MS Certified
Sample Vials W hitepaper,
Literature Reference 720001517EN
www.waters.com
Vials are available in three closure types: crimp, snap, and screw cap. Each
closure has its advantages and disadvantages.
Crimp caps squeeze the septum between the rim of the glass vial and the
crimped aluminum cap. T his forms an excellent seal preventing evaporation.
T he septum stays seated during piercing by the autosampler needle. T he
crimp cap vial requires crimping tools to carry out the sealing process. For
few samples, manual crimper tools are the best choice. For large numbers of
sample, automated crimpers are available.
Snap caps are an extension of the crimp cap system of sealing. A plastic
cap is stretc hed over the rim of the vial to form a seal by squeezing the
septum between the glass and the stretc hed plastic cap. T he plastic cap
creates tension when trying to return to its original size. T his tension forms
the seal between glass, cap and septum. Plastic snap caps do not require
any tools to assemble.
Snap caps are not as effective a seal as other closures.
Screw caps are universal. Screwing the cap applies a mechanical force that
squeezes the septum between the glass rim and the cap. Screw caps form
an excellent seal and mechanically hold the septum in place during piercing.
No tools are required for assembly.
LectraBond screw caps are available through Waters. T his screw cap
has a PT FE/silicone septum bonded to the polypropylene cap, using a
non-solvent bonding process. T his bonding tec hnology is designed to
keep the septum/cap together during shipment and assembly onto vials.
T he bond will aid in preventing dislodging of the septum during use,
but the primary sealing mec hanism is the mec hanical force applied by
tightening the cap to the vial.
Cap tightening is the mechanism that forms the seal and holds the septum
in place during needle insertion. T here is no need to over-tighten the cap,
as it can compromise the seal and lead to dislodging. T he septum starts to
cup or indent when you begin to over-tighten.
Cap Design
Strength Design
Comment
Crimp
Excellent seal
Requires tools
Snap
Moderate seal
Screw
Excellent seal
Universal
www.waters.com
63
Amber
Max Recovery
Amber Max
300 L PP
750 L PP
Clear Glass
with Septumless Cap
600000668CV
600000669CV
600000670CV
600000755CV
186003886C
186000307C
186000847C
186000327C
186000307DV
186000847DV
186000327DV
186004132C
186004133C
186004168C
186004132C
Combination Packs
Bonded Pre-slit Silicone/PTFE Septum
186002639
186004112
186005221
1600 L
1600 L
1100 L
1100 L
210 L
530 L
1600 L
Residual*
165 L
165 L
22 L
22 L
20 L
70 L
165 L
* If you need to access more sample, go to the advanced setting screen in the Sample Manager Instrument Method Editor to change the needle placement.
For the Amino Acid ACQUITY UPLC System and the MassTrack Amino Acid Analysis Solution System, use of Waters Total Recovery Vials are recommended, Part Number 186004631.
Part No.
405003743
Clear 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with Quick Thread Design,
(6 mm Opening, 9 mm Cap).
Amber 12 x 32, Type 1, 51-Expansion Glass Screw Neck with Quick Thread Design,
(6 mm Opening, 9 mm Cap).
Waters Maximum Recovery Vial, 12 x 32, Type 1, 33-Expansion Glass, Screw Neck
with Quick Thread Design (6 mm Opening, 9 mm Cap).
Clear 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with Quick Thread Design,
(6 mm Opening, 9 mm Septumless Cap).
www.waters.com/vialsselector
T his interactive tool requires that the user has Adobe Flash Player 8.
64
www.waters.com
Description
Residual Volume
Needle Placement
Comments
1,2,7
38.3
32.0
165 L
2 mm
Preset
1,2,7
38.3
31.0
4 L
3 mm
Advanced Settings
3,4
38.3
32.0
22 L
2 mm
Preset
300 PP Vial
38.3
31.0
20 L
3 mm
Advanced Settings
750 PP Vial
38.3
31.5
70 L
3 mm
Advanced Settings
*Please reference the listing of Vials Descriptions on pages 78 and 79 for more details.
The table above shows vial depths, residual volumes and needle placements
for several products. In the ACQUITY UPLC system, the default needle
placement for 48-well vial holders with a 2 mL vial (ANSI 48 vial 2 ml
holder) is automatically preset and brings the needle down to a depth
which is 2 mm from the bottom of the vial. T he residual sample volume
remaining in the vial for a 2 mm offset is recorded in the table above.
For situations where you need to access more of the sample, or if you need
to keep the needle further from the bottom of the vial, go to the advanced
setting screen in the Sample Manager Instrument Method Editor to change
the needle placement.
Please note that you must go to the advanced settings and change the
needle placement to 3 mm for vial number 5 in the table above. You will
also have to review needle settings when using glass inserts.
Waters ACQUITY UPLC System Sample Manager Version 1.30 and later
Sample Needle
Vial Depth
Typical 2 mL Vial
2 mm Default Offset
www.waters.com
65
384-Well Plates
186002643
186002481
186002482
186002632
100
50
50
50
50
350 L
800 L
2 mL
250 L
100 L
186002483
186002483
186002484
186002788
186002788
186002788
186002788
186002788
186002789
186002789
186002789
186002789
186002789
35 L
15 L
20 L
15 L
15 L
Part No.
Max Volume
Residual Volume
186001437 (DV)2
650 L
15 L
850 L
15 L
186000349 (DV)2
650 L
15 L
186000855 (DV)2
850 L
15 L
Pack Size
Well Volume
186002631
Sealing Options
* If you need to access more sample, go to the advanced setting screen in the Sample Manager Instrument Method Editor to change the needle placement.
1
2
1860014381
186001436 (DV)
Sealing Cap for 700 L Glass Insert Square Well Seals Against the Well Wall PTFE/Silicone, 5/pk
186000857
Sealing Cap for 1 mL Glass Inserts Seals in the Glass PTFE/Silicone, 10/pk
186000856
Quick-load packs are the easy and fast way to load your plate with glass inserts
66
www.waters.com
Part No.
115 Volt
186002786
240 Volt
186002787
Temperature Range
Solvent Range
Clear Polyester
From -80 C to 80 C
From -200 C to 90 C
Recommendation
Best for Long Term Storage
Literature
References
Sample Vials and Accessories Brochure,
Literature Reference 720001818EN
Waters Heat Sealer User Manual,
Literature Reference 720001330EN
96-well Collection Plate Options for the
Waters Extraction Plate Manifold,
Literature Reference 720001263EN
Waters 96 and 384-Well
Collection Plate Specifications,
Literature Reference WA41941
Waters LC/MS Certified
Sample Vials W hitepaper,
Literature Reference 720001517EN
Determination of the Level of Ion
Suppression from LC/MS Vials,
Literature Reference WA60004
Waters Certified Sample Vials
Technical W hitepaper,
Literature Reference 720001303EN
www.waters.com
67
Clear
Amber
Max Recovery
300 L PP
10 mm Cap Clear
Total Recovery
Amber Max
10
11
12
13
14
15
600000668CV
600000669CV
600000670CV
600000671CV
600000755CV
WAT270946C
186004132C
Combination Packs
Bonded Pre-slit Silicone/PTFE Septum
186000307C
186000847C
186000327C
Combination Deactivated
186000307DV
186000847DV
186000327DV
186002639*
186000385C
186003886C
Max
1100 L
1100 L
280 L
1100 L
950 L
1100 L
Residual
750 L
750 L
20 L
750 L
9 L
750 L
Max
1700 L
1700 L
1500 L
290 L
1700 L
1500 L
1700 L
Residual
170 L
170 L
22 L
10 L
170 L
22 L
170 L
186000385DV
Insert
WAT094171 (DV)1 WAT094171 (DV)1
144 L/6 L
144 L/6 L
Alliance 2690/2695
Alliance 2790/2795
WAT094171 (DV)1
144 L/6 L
144 L/6 L
Compatible Systems
When (DV) appears beside the part number, a deactivated version of this product can be ordered by adding DV to the right of the part number.
* Not certified
For more details, see vials descriptions on pages 78 and 79.
68
www.waters.com
384-Well Plates
186002643
186002481
186002482
186002632
100
50
50
50
50
350 L
800 L
2 mL
250 L
100 L
186002483
186002483
186002484
186002788
186002788
186002788
186002788
186002788
186002789
186002789
186002789
186002789
186002789
35 L
15 L
20 L
Pack Size
Well Volume
186002631
Sealing Options
Heat Sealer
Description
Description
Part No.
115 Volt
186002786
240 Volt
186002787
Part No.
Part No.
186002633
1860014381
186001437(DV)2
186001436(DV)2
186000349(DV)2
186000855(DV)2
186000857
Sealing Cap for 1 mL Round Well Seals in the Glass PTFE/Silicone, 10/pk
186000856
Heat sealer
www.waters.com/plateselector
T his interactive tool requires that the user has Adobe Flash Player 8.
www.waters.com
69
Amber
Max Recovery
300 L PP
750 L PP
10 mm Cap Clear
Total Recovery
Amber Max
16
17
18
19
20
21
22
23
600000751CV
600000752CV
600000749CV
600000750CV
600000754CV
600000668CV
600000669CV
600000670CV
600000671CV
600000755CV
186000272C
186000846C
186000326C
WAT270946C 2
186000384C
186003885C
Combination Deactivated
186000272DV
186000846DV
186000326DV
WAT270946DV 2
186000384DV
186000307C
186000847C
186000327C
Combination Deactivated
186000307DV
186000847DV
186000327DV
186004132C
186004133C
186004168C
1860041123
186002802
186002626
1860026403
1860026393
1860052203
1860052213
186000385C
186003886C
186000385DV
186004167C
Vials Only
Vials Only
186000273
186000848
186000273DV
186000848DV
Max
1100 L
1100 L
280 L
400 L
1100 L
950 L
Residual
750 L
750 L
20 L
300 L
750 L
9 L
Max
1700 L
1700 L
1500 L
290 L
530 L
1700 L
1500 L
Residual
170 L
170 L
22 L
10 L
170 L
170 L
22 L
186005219
WAT063300
186002805
WAT063300DV
Inserts
WAT094170(DV)1 WAT094170(DV)1
230 L/20 L
WAT094170(DV)1
230 L/20 L
230 L/20 L
WAT094171(DV)1 WAT094171(DV)1
144 L/6 L
144 L/6 L
186004169
186004169
WAT094171(DV)1
144 L/6 L
186004169
186004169
186004169
Cap Black
WAT058875
Septa Silicone/PTFE
WAT058874
186004169
186004169
186004169
186004169
186004169
186000274
186004169
Blue LectraBond
186000274
186000274
186000274
186000274
186002129
186000274
Red LectraBond
186002129
186002129
186002129
186002129
186002130
186002129
186002456
Green LectraBond
186002130
186002130
186002130
186002130
White LectraBond
186002456
186002456
186002456
186002456
186002130
Blue LectraBond
186000305
186000305
186000305
186000305
186000305
186000305
Red LectraBond
186002128
186002128
186002128
186002128
186002128
186002128
Green LectraBond
186002127
186002127
186002127
186002127
186002127
186002127
White LectraBond
186002457
186002457
186002457
186002457
186002457
186002457
186000989(DV)1
186003455
Alliance 2690/2695
Alliance 2790/2795
186002456
70
When (DV) appears beside the part number, a deactivated version of this product can be ordered by adding DV to the right of the part number.
www.waters.com
Amber
Max Recovery
PP 300 L
PP 750 L
Amber Crimp
Total Recovery
24
25
26
27
28
29
30
31
186002642
186005223
186002641
186005222
186002628
186005224
Combination Packs
186000234(DV)1
Vials
Vials Only
WAT094219
WAT094220
186000984
WAT094219DV
WAT094220DV
186000984DV
Max
1100 L
1100 L
280 L
Residual
750 L
750 L
20 L
Max
1700 L
1700 L
1500 L
Residual
170 L
170 L
22 L
WAT094222
WAT094223
186000302
WAT094222DV
WAT094223DV
186000302DV
400 L
1100 L
1100 L
950 L
300 L
750 L
750 L
9 L
290 L
530 L
1700 L
1700 L
10 L
170 L
170 L
170 L
Inserts
300 L with Poly Spring
Max Volume Injection/Max Residual Volume
150 L with Poly Spring
Max Volume Injection/Max Residual Volume
WAT094170(DV)1 WAT094170(DV)1
230 L/20 L
230 L/20 L
230 L/20 L
WAT094171(DV)1 WAT094171(DV)1
144 L/6 L
144 L/6 L
144 L/6 L
144 L/6 L
Blue
186000303
186000303
186000303
186000303
186000303
Black
186002649
186002649
186002649
186002649
186002649
186002649
Red
186002650
186002650
186002650
186002650
186002650
186002650
Blue
186000304
186000304
186000304
186000304
186000304
186000304
Black
186002648
186002648
186002648
186002648
186002648
186002648
Red
186002647
186002647
186002647
186002647
186002647
186002647
Blue
186000328
186000328
186000328
186000328
186000328
186000328
Black
186002645
186002645
186002645
186002645
186002645
186002645
Red
186002646
186002646
186002646
186002646
186002646
186002646
Crimp Cap
Crimp Cap Silicone/PTFE Septum
PSL404219
PSL404219
PSL404231
PSL404231
Compatible Systems
Alliance 2690/2695
Alliance 2790/2795
When (DV) appears beside the part number, a deactivated version of this
product can be ordered by adding DV to the right of the part number.
Part No.
186004487
www.waters.com
71
Total Recovery
PP Conical
Amber Glass
Shell Vial
32
33
34
35
36
37
38
186002629C
186004031
WAT025051
WAT025050
2950 L/50 L
2400 L/1600 L
2400 L/1600 L
15 x 45 mm Vials
48 Position Carousel
Combination Packs
Vial, Cap and LectraBond PTFE/Silicone Septum
186000838C
186001133C
Combination Deactivated
186000838DV
186001133DV
186000839C
186001134C
Combination Deactivated
186000839DV
186001134DV
186002630C
186000840(DV)1
186001135(DV)1
186002520
1860009992
2400 L/1600 L
2400 L/1600 L
3000 L/40 L
2000 L/1000 L
186000841
186000841
186000841
186000841
186000965
186000965
186000965
186000965
186000842
186000842
186000842
186000842
WAT072711
WAT072711
WAT072711
WAT072711
WAT073005
WAT073005
WAT073005
WAT073005
WAT072714
WAT072714
WAT072714
WAT072714
WAT022861
WAT022861
WAT022861
WAT022861
WAT072704(DV)1
WAT072704(DV)1
244 L/6 L
244 L/6 L
WAT015199(DV)1
WAT015199(DV)1
230 L/20 L
230 L/20 L
WAT072030
WAT072030
230 L/20 L
230 L/20 L
WAT072708
WAT072708
186001729
186001729
WAT072704
When (DV) appears beside the part number, a deactivated version of this product can be ordered by adding DV to the right of the part number.
2 Item contains 1,000 vials
3 Inserts requires springs (Part No. WAT072708)
1 mL Shell
Amber
Total Recovery
PP Conical
39
40
41
42
8 x 40 mm Vials
96 Position Carousel
WAT025054C
WAT025053C
186000837C
WAT025054DV
WAT025053DV
186000837DV
Pack Size
Max Volume Injection/Max Residual Volume
150 L Glass Insert
WAT022476
250
250
100
100
600 L/400 L
600 L/400 L
700 L/6 L
650 L/50 L
WAT072294(DV)1
WAT072294(DV)1
WAT072289
WAT078515
WAT078515
WAT072289
144 L/6 L
144 L/6 L
WAT078515
WAT078515
186001728
186001728
WAT022479
When (DV) appears beside the part number, a deactivated version of this
product can be ordered by adding DV to the right of the part number.
72
www.waters.com
Amber
Max Recovery
Amber Max
300 L PP
43
44
45
46
47
600000668CV
600000669CV
600000670CV
600000755CV
186000307C
186000847C
186000327C
186003886C
186000307DV
186000847DV
186000327DV
186000272C
186000846C
186000326C
186003885C
Combination Packs
Bonded Pre-slit Silicone/PTFE Septum
186002639
186002640
1600 L
1600 L
1100 L
1100 L
240 L
Residual
150 L
150 L
10 L
10 L
10 L
WAT094171
WAT094171
144 L/6 L
144 L/6 L
Components
384-Well Plates
186002643
186002481
186002482
186002632
100
50
50
50
50
350 L
800 L
2 mL
250 L
100 L
186002483
186002483
186002484
186002788
186002788
186002788
186002788
186002788
186002789
186002789
186002789
186002789
186002789
125 L
40 L
60 L
40 L
40 L
Part No.
Max Volume
Residual Volume
15 L
Pack Size
Well Volume
186002631
Sealing Options
1860014381
186001437 (DV)2
650 L
186001436 (DV)2
850 L
15 L
186000349 (DV)2
650 L
15 L
186000855 (DV)2
850 L
15 L
Sealing Cap for 700 L Glass Insert Square Well Seals Against the Well Wall PTFE/Silicone, 5/pk
186000857
Sealing Cap for 1 mL Glass Inserts Seals in the Glass PTFE/Silicone, 10/pk
186000856
When (DV) appears beside the part number, a deactivated version of this product can be ordered by adding DV to the right of the part number.
Components
22 x 45 mm Clear Glass Vial
186001420
186004632
www.waters.com
48
73
Amber
Amber Max
Recovery
Clear Glass
Max Recovery
Qsert Vial
Amber Qsert
PP 300 L
PP 750 L
10 mm Cap
PP 250 L
8 mm Cap
49
50
51
52
53
54
55
56
57
58
186000326C
186001126C
186001130C
WAT270946C1
186000326DV
186001126DV
186001130DV
WAT270946DV1
186000327C
186001128C
186001131C
186000327DV
186001128DV
186001131DV
600000751CV
186000272C
186000846C
Combination Deactivated
186000272DV 186000846DV
Bonded Pre-slit
Silicone/PTFE Septum
186000307C
Combination Deactivated
186000307DV 186000847DV
186000847C
186003885C
186003886C
Combination Packs
Bonded Silicone/PTFE Septum
186002640
186005220
186002639
186005221
186002626
186005219
Vials Only
186000273
Vials Only
Deactivated Vials Only
186000848
186002802
186002804
186002803
186000273DV 186000848DV
WAT063300
WAT094172
WAT063300DV
Inserts
300 L with Poly Spring
300 L with Poly Spring Deactivated
150 L with Poly Spring
150 L with Poly Spring Deactivated
WAT094170
WAT094170
WAT094170
WAT094170DV
WAT094170DV
WAT094170DV
WAT094171
WAT094171
WAT094171
WAT094171DV
WAT094171DV
WAT094171DV
186004169
186004169
186004169
186004169
186004169
186004169
186004169
Black Cap
WAT058875
WAT094174
WAT058876
WAT058874
WAT210685
WAT058886
Blue LectraBond
186000274
186000274
186000274
186000274
186000274
186000274
Red LectraBond
186002129
186002129
186002129
186002129
186002129
186002129
186002129
Green LectraBond
186002130
186002130
186002130
186002130
186002130
186002130
186002130
186000305
186000305
186000305
186000305
186000305
186000305
186000305
Red LectraBond
186002128
186002128
186002128
186002128
186002128
186002128
186002128
Green LectraBond
186002127
186002127
186002127
186002127
186002127
186002127
186002127
Compatible Systems
Agilent Technologies
Alcott, Antek, CTC, Spark
Thermal Separations
Beckman, Dynatech, Finnigan,
Fisons, Gilson
74
www.waters.com
Amber
Max Recovery
Qsert Vial
PP 300 L
PP 750 L
Clear Crimp
Amber
Crimp
59
60
61
62
63
64
65
66
186001124(DV)1
186002642
186005223
186002641
186005222
186002628
186005224
Combination Packs
Vial, Cap and Silicone/PTFE Septum
Vial, Cap and Pre-Slit
Silicone/PTFE Septum
186001125(DV)
186001127(DV)
Vials Only
Vials Only
WAT094219
WAT094220
186000984
WAT094219DV
WAT094220DV
186000984DV
WAT094222
WAT094223
WAT094222DV
WAT094223DV
WAT094170(DV)1
WAT094171(DV)1
WAT094170(DV)1
WAT094170(DV)1
WAT094170(DV)1
WAT094171(DV)1
WAT094171(DV)1
WAT094171(DV)1
Blue
186000303
186000303
186000303
186000303
186000303
Black
186002649
186002649
186002649
186002649
186002649
186002649
Red
186002650
186002650
186002650
186002650
186002650
186002650
PSL404219
PSL404219
PSL404231
PSL404231
186000304
186000304
186000304
186000304
186000304
186000304
Black
186002648
186002648
186002648
186002648
186002648
186002648
Red
186002647
186002647
186002647
186002647
186002647
186002647
Blue
186000328
186000328
186000328
186000328
186000328
186000328
Black
186002645
186002645
186002645
186002645
186002645
186002645
Red
186002646
186002646
186002646
186002646
186002646
186002646
Crimp Cap
Compatible Systems
Agilent Technologies, Beckman,
Dynatech, Finnigan, Fisons,
Gilson, Hitachi, LDC, Perkin-Elmer,
Shimadzu, Spectra-Physics, Varian
When (DV) appears beside the part number, a deactivated version of this product can be ordered by adding DV to the right of the part number.
Part No.
186004487
www.waters.com
75
Amber
Screw Neck
Total Recovery
PP Screw
Neck Vial
PP Snap Cap
Amber Glass
Shell Vial
67
68
69
70
71
72
73
186002629C
186004031
WAT025051
WAT025050
Combination Pack
Vial, Cap and LectraBond PTFE/Silicone Septum
186000838C
186001133C
Combination Deactivated
186000838DV
186001133DV
186000839C
186001134C
Combination Deactivated
186000839DV
186001134DV
186002630C
186000840
186001135
186000840DV
186001135DV
1860009991
186002520
186000841
186000841
186000841
186000965
186000965
186000965
186000842
186000842
186000842
186000965
WAT072711
WAT072711
WAT072711
WAT073005
WAT073005
WAT073005
WAT072714
WAT072714
WAT072714
WAT022861
WAT022861
WAT022861
WAT072704
WAT072704
WAT072704
WAT072704DV
WAT072704DV
WAT072704DV
WAT015199
WAT015199
WAT015199
WAT015199DV
WAT015199DV
WAT015199DV
WAT072030
WAT072030
WAT072030
WAT072708
WAT072708
WAT072708
186001729
186001729
186001729
4 mL Screw Cap
10 mL Screw Neck
Compatible Systems
Bruker, Kontron, Perkin-Elmer, Shimadzu, Tosoh, Unicam
1
75
Vial
186000840
186001420
600000162
186001421
PTFE Septum
WAT0727141
186001422
Components
76
186004108
186004109
186004111
www.waters.com
Average Thickness
Needle Offset
Notes*
Comments
Description
1,3
1,3
11
1 mm
27
1 mm
22
31
21
1,3
24
1,3
29
1 mm
2,3
58
24
29
21
21
1 mm
1 mm
29
30
Vial (6 mm neck)
30
30
Vial (6 mm neck)
30
Snap/Crimp Vial
1 mm
2,3
15
1,3
1 mm
*Notes:
1. Clears needle tipno offset required. Meets the criteria of a bottom thickness < 1.6 mm.
2. Does not clear needle tippositive needle offset should be used.
3. Designed to accept Waters low volume inserts (LVIs)at least 1 mm offset should be added.
4. T his dimension should be added to vial bottom thickness and result checked against the criteria for bottom thickness.
Literature
References
Sample Vials & Accessories Brochure,
Literature Reference 720001818EN
Waters LC/MS Certified
Sample Vials W hitepaper,
Literature Reference 720001517EN
www.waters.com
77
Clear 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with Quick Thread Design
(6 mm Opening, 9 mm Cap).
32
Amber 12 x 32, Type 1, 51-Expansion Glass Screw Neck with Quick Thread Design
(6 mm Opening, 9 mm Cap).
33
Clear Maximum Recovery Vial 12 x 32, Type 1 33-Expansion Glass, Screw Neck with
Quick Thread Design (6 mm Opening, 9 mm Cap).
34
Total Recovery Screw Neck Vial Clear Glass 15 x 45, Type 1, 33-Expansion Glass.
10
35
11
Polypropylene 12 x 32, 300 L Screw Neck with Quick Thread Design (6 mm Opening,
9 mm Cap). Reformulate Clean PP Vial.
36
12
37
4 mL Glass Shell Vial with Polyethylene Snap Cap, Type 1, 51-Expansion Glass.
13
Total Recovery Vial Clear 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with
Quick Thread Design (6 mm Opening, 9 mm Cap).
38
4 mL Amber Shell Vial with Polyethylene Snap Cap, Type 1, 51-Expansion Glass.
14
Amber Maximum Recovery Vial, 12 x 32, Type 1 51-Expansion Glass, Screw Neck with
Quick Thread Design (6 mm Opening, 9 mm Cap).
39
15
Clear 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with Quick Thread Design,
(6 mm Opening, 9 mm Septumless Cap).
40
41
Total Recovery Clear Glass Vial (8 x 40 mm) with Polyethylene Snap Cap,
Type 1, 51-Expansion Glass.
42
16
Clear 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with Quick Thread Design
(6 mm Opening, 9 mm Cap).
17
Amber 12 x 32, Type 1, 51-Expansion Glass Screw Neck with Quick Thread Design
(6 mm Opening, 9 mm Cap).
18
Clear Maximum Recovery Vial 12 x 32, Type 1 33-Expansion Glass, Screw Neck with
Quick Thread Design (6 mm Opening, 9 mm Cap).
43
Clear 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with Quick Thread Design,
(6 mm Opening, 9 mm Cap).
19
Polypropylene 12 x 32, 300 L Screw Neck with Quick Thread Design (6 mm Opening,
9 mm Cap). Reformulate Clean PP Vial.
44
20
Polypropylene 12 x 32, 750 L Screw Neck with Quick Thread Design (6 mm Opening,
9 mm Cap). Reformulate Clean PP Vial.
Amber 12 x 32, Type 1, 51-Expansion Glass Screw Neck with Quick Thread Design,
(6 mm Opening, 9 mm Cap).
45
Waters Maximum Recovery Vial, 12 x 32, Type 1, 33-Expansion Glass, Screw Neck
with Quick Thread Design (6 mm Opening, 9 mm Cap).
46
47
48
21
22
Total Recovery Vial Clear 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with
Quick Thread Design (6 mm Opening, 9 mm Cap).
23
Amber Maximum Recovery Vial, 12 x 32, Type 1 51-Expansion Glass, Screw Neck with
Quick Thread Design (6 mm Opening, 9 mm Cap).
24
Clear 12 x 32, Type 1, 33-Expansion Glass, Snap Cap Vial (6 mm Opening, 9 mm Cap).
25
Amber 12 x 32, Type 1, 51-Expansion Glass Snap Cap Vial (6 mm Ppening, 9 mm Cap).
26
Clear Maximum Recovery Vial 12 x 32, Type 1, 33-Expansion Glass, Snap Cap Vial
(6 mm Opening, 9 mm Cap).
49
Clear 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with Quick Thread Design
(6 mm Opening, 9 mm Cap).
27
50
Amber 12 x 32, Type 1, 51-Expansion Glass Screw Neck with Quick Thread Design
(6 mm Opening, 9 mm Cap).
28
51
Amber Maximum Recovery Vial 12 x 32, Type 1, 51-Expansion Glass, Screw Neck with
Quick Thread Design (6 mm Opening, 9 mm Cap).
29
Clear 12 x 32, Type 1, 33-Expansion Glass, Crimp Top Vial (6 mm Opening, 12 mm Cap).
52
30
Amber 12 x 32, Type 1, 51-Expansion Glass, Crimp Top Vial (6 mm Opening, 12 mm Cap).
Clear Maximum Recovery Vial 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with
Quick Thread Design (6 mm Opening, 9 mm Cap).
53
31
Total Recovery Vial Clear 12 x 32, Type 1, 33-Expansion Glass, Snap Cap Vial
(6 mm Opening, 9 mm Cap).
Qsert Vial Clear Screw Cap Glass Vial, Quick Thread Design with Fused in Glass Insert
(6 mm Opening, 9 mm Cap).
54
Qsert Vial Amber Screw Cap Glass Vial, Quick Thread Design with Fused in Glass Insert
(6 mm Opening, 9 mm Cap).
55
Polypropylene 12 x 32, 300 L Screw Neck with Quick Thread Design (6 mm Opening,
9 mm Cap). Reformulate Clean PP Vial.
56
Polypropylene 12 x 32, 750 L Screw Neck with Quick Thread Design (6 mm Opening,
9 mm Cap). Reformulate Clean PP Vial.
57
58
78
www.waters.com
Clear 12 x 32, Type 1, 33-Expansion Glass, Snap Cap Vial (6 mm Opening, 9 mm Cap).
67
60
Amber 12 x 32, Type 1, 51-Expansion Glass Snap Cap Vial (6 mm Opening, 9 mm Cap).
68
61
Maximum Recovery Vial 12 x 32, Type 1, 33-Expansion Glass, Snap Cap Vial
6 mm Opening, 9 mm Cap).
69
Waters Total Recovery Screw Neck Vial Clear Glass 15 x 45, Type 1, 33-Expansion Glass.
62
Qsert Vial Clear Snap Cap Glass Vial with Fused in Glass Insert (6 mm Opening,
9 mm Cap).
70
71
63
72
4 mL Glass Shell Vial with Polyethylene Snap Cap, Type 1, 51-Expansion Glass.
64
73
4 mL Amber Shell Vial with Polyethylene Snap Cap, Type 1, 51-Expansion Glass.
65
Clear 12 x 32, Type 1, 33-Expansion Glass, Crimp Top Vial (6 mm Opening, 12 mm Cap).
66
Amber 12 x 32, Type 1, 51-Expansion Glass, Crimp Top Vial (6 mm Opening, 12 mm Cap).
4 mL Glass Screw Neck Vial, Type 1, 33-Expansion Glass for GPC 2000.
75
Impact
Solution
The glass inserts in the Waters 96-well format allows for the
use of a sealing cap mat, saving time and labor.
Waters vials are distributed world wide from the same source.
Only Waters Alliance Total Recovery vials and Maximum Recovery vials utilize a proprietary manufacturing process,
ensuring that the slope of the internal taper will deliver all of the sample to the bottom of the vial.
T he bottom thickness is held to a close tolerance eliminating needle damage caused by bottoming out.
www.waters.com
79
FILTRaTION
3. Add plasma.
Ion suppression
MS extractables
Ion enhancement
Recovery of analytes
UV extractables
80
Filtration
Description
Qty.
Part No.
1/pk
5/pk
186003873
186002448
www.waters.com
FILTRaTION
Nature
of
Sample
Aqueous
GHP* universal
Nylon
PVDF
PES
Ion
Chromatography
IC
Non Polar
CR/
PTFE
GHP
Protein
GHP
GH
PVDF
For samples with laden particulate that are difficult to filter, it is best to use a syringe filter with a glass fiber
pre-filter over the membrane. These are available in GHP and Nylon.
> 3 m
0.45 m
< 3 m
0.20 m
Membrane Choices
GHP Acrodiscs
Volume
Acrodisc Size
Nylon Acrodiscs
< 2 mL
4 mm
< 10 L
< 10 mL
13 mm minispike
< 14 L
< 10 mL
13 mm male luer
< 30 L
< 100 mL
25 mm
< 100 L
Hold Up Volume
Acrodisc LC (PVDF)
Question
Answer
Choice
Sample
Aqueous
5 m
0.45 m
Volume
1.5 mL
4 mm or larger
Acrodisc CR(PTFE)
www.waters.com
Filtration
81
FILTRaTION
Acetone
Acetoniltrile
n-Butanol
Chloroform
Dioxane
Dimethyl formamide
Dimethyl sulfoxide
Ethanol
Ethyl acetate
Ethyl ether
Freon TF
Hexane, dry
Methanol
Methylene chloride
N-Methylpyrrolidone
Isopropanol
Tetrahydrofuran
Tetrahydrofuran/
water (50/50)
Toluene
Water
R
LR
NR
= RESISTANT
= LIMITED RESISTANCE
= NOT RESISTANT
= INSUFFICIENT DATA
R*
LR
R*
R*
R*
LR
R*
R*
LR
LR
LR*
R*
R*
R*
R*
R*
R*
R*
R*
R*
R*
R*
R*
R*
R*
NR*
NR*
NR*
R*
NR*
NR*
NR
R*
NR*
NR*
NR*
R*
NR*
NR*
NR
R*
R*
NR
R*
R*
R*
NR
NR
NR
R*
R*
NR
LR
R*
R*
NR
R*
R*
R*
NR
NR
NR
R*
R*
NR
LR
R*
R*
NR
R*
R*
R*
NR
NR
NR
R*
R*
NR
LR
R*
R*
NR
R*
R*
R*
NR
NR
NR
R*
R*
NR
LR
R*
NR
LR
NR
NR
NR
NR
LR
LR
LR
NR
NR
NR
LR
NR
NR
NR
NR
NR
NR
NR
NR
NR
LR
NR
NR
NR
NR
NR
NR
NR
GH Polypro
FP Verciel (PVDF)
NR
LR
NR
NR
LR
NR
NR
LR
Nylaflo (Nylon)
NR
NR
LR
NR
LR
NR
NR
NR
TF (PTFE)
Disc Filters
Note:
R = Resistant
No significant change was observed in flow rate or bubble point of the membrane.
4 mm Polypropylene
Housing Male Luer
13 mm Polypropylene
Housing Male Luer
82
Filtration
13 mm Polypropylene
Housing Minispike
25 mm Polypropylene
Housing Male Luer
www.waters.com
FILTRATION
Part Number Cross Reference
Pall Life Sciences
Description
PTFE, 4 mm, 0.45 m
PTFE, 13 mm, 0.2 m
PTFE, 13 mm, 0.45 m
PTFE, 25 mm, 0.2 m
PTFE, 25 mm, 0.45 m
PVDF, 13 mm, 0.45 m
PVDF, 25 mm, 0.45 m
GHP, 13 mm, 0.45 m
GHP, 25 mm, 0.45 m
Nylon, 13 mm, 0.2 m
Nylon, 13 mm, 0.45m
Nylon, 25 mm, 0.2 m
Nylon, 25 mm, 0.45 m
Supor, 25 mm, 0.2 m
PVDF, 47 mm, 0.45 m
PVDF, 47 mm, 0.2 m
Nylon, 47 mm, 0.45 m
Nylon, 47 mm, 0.2 m
PTFE, 47 mm, 0.45 m
PTFE, 47 mm, 0.2 m
GHP, 47 mm, 0.45 m
Supor, 47 mm, 0.2 m
Supor, 47 mm, 0.45 m
Supor, 13 mm, 0.45 m
Glass Filter Holder
Forceps
300 mL Funnel
Glass Base, Tabulated Cap
Ground Joint Flask
Thick Glass Prefilter
AVE Glass Prefilter
Whatman
Part No.
Millipore
Part No.
Waters
Part No.
6783-0404
6783-1302
6783-1304
6784-2502
6784-2504
6792-1304
6784-1304
6785-2504
6790-1302
6790-1304
6780-2502
7404-004
7402-004
7585-004
7582-004
7002-0447
SJFHL04NS
SJFG013NS
SJFH013NS
SLFG025NS
SLSR025NS
SJHV013NS
SLHV025NS
SJFH013NS
SLCR025NS
SLGS025OS
HVLP04700
FHLP04700
FGLP04700
FHLC04700
GSTF04700
HATF04700
HATF01300
XX1504700
XX6200006
XX1004704
XX1504702
XX1504705
AP2501000
AP4001000
WAT200508
WAT200506
WAT200502
WAT200504
WAT200500
WAT200512
WAT200510
WAT200516
WAT200514
WAT200524
WAT200520
WAT200522
WAT200518
WAT200529
WAT200530
WAT200531
WAT200532
WAT200533
WAT200534
WAT200535
WAT200537
WAT200539
WAT200538
WAT200540
WAT200543
WAT200544
WAT200545
WAT200546
WAT200547
WAT200541
WAT200542
Description
Qty.
Diameter
Pore Size
Part No.
PVDF Filter
Nylon Filter
PTFE Filter
100/pk
100/pk
100/pk
100/pk
100/pk
100/pk
100/pk
100/pk
100/pk
100/pk
100/pk
100/pk
100/pk
100/pk
47 mm
47 mm
47 mm
13 mm
47 mm
47 mm
13 mm
47 mm
47 mm
47 mm
47 mm
47 mm
10 mm
10 mm
0.45 m
0.45 m
0.45 m
0.45 m
0.45 m
0.45 m
0.45 m
0.2 m
0.2 m
0.2 m
0.2 mm
0.2 m
1.0 m
1.0 m
WAT200530
WAT200532
WAT200534
WAT200536
WAT200537
WAT200538
WAT200540
WAT200531
WAT200533
WAT200535
186003524
WAT200539
WAT200541
WAT200542
GH Polypro Filter
Supor (PES) Filter
PVDF Filter
Nylon Filter
PTFE Filter
GHP
Supor (PES) Filter
Thick Glass Filter
A/E Glass Filter
Funnel, 300 mL
WAT200545
Glass Base,
tabulated cap
WAT200546
Ground
Joint Flask
WAT200547
Description
www.waters.com/filterselector
Part No.
WAT085113
WAT085102
WAT200543
WAT200544
WAT200545
WAT200546
WAT200547
WAT200566
WAT085114
WAT085123
WAT085115
T his interactive tool requires that the user has Adobe Flash Player 8.
www.waters.com
Filtration
83
FILTRaTION
Syringe Filters Product List
Acrodisc 13 mm
Pack Size
100
0.2 m
300
1000
100
0.45 m
300
1000
Aqueous
NYLON
PVDF
WAT200524
WAT200806
WAT200525
WAT200807
WAT200834
WAT200520
WAT200512
WAT200521
WAT200513
WAT200832
WAT200827
Non Polar
CR
WAT200506
WAT200507
WAT200823
WAT200502
WAT200503
WAT200821
Protein
PVDF
WAT200806
WAT200807
WAT200512
WAT200513
WAT200827
Ion Chromatography
IC
WAT200810
WAT200811
WAT200844
WAT200812
WAT200813
WAT200842
100
0.2 m
300
1000
100
0.45 m
300
1000
GHP
NYLON
PVDF
WAT097962
WAT200562
WAT200804
WAT097963
WAT200563
WAT200805
186004567
WAT200835
WAT200838
WAT200516
WAT200564
WAT200560
WAT200517
WAT200565
WAT200561
WAT200830
WAT200836
WAT200828
Non Polar
CR
GHP
WAT200556
WAT097962
WAT200557
WAT097963
WAT200824
186004567
WAT200558
WAT200516
WAT200559
WAT200517
WAT200825
WAT200830
Protein
PVDF
WAT200804
WAT200805
WAT200838
WAT200560
WAT200561
WAT200828
50
0.2 m
200
1000
50
0.45 m
200
1000
Acrodisc 25 mm
Pack Size
Aqueous
GHP
NYLON
PVDF
GHP GF*
NYLON GF*
GF**
Versapor
Sterile Syringe PES
WAT097964
WAT200522
WAT200808
WAT200529
WAT097965
WAT200523
WAT200809
186004566
WAT200833
WAT200839
WAT200514
WAT200518
WAT200510
WAT200802
WAT200800
WAT200818
WAT200515
WAT200519
WAT200511
WAT200803
WAT200801
WAT200819
WAT200829
WAT200831
WAT200826
WAT058853
WAT200846
WAT200840
WAT200841
Non Polar
CR
GHP
WAT200504
WAT097964
WAT200505
WAT097965
WAT200822
186004566
WAT200500
WAT200514
WAT200501
WAT200515
WAT200820
WAT200829
Protein
PVDF
WAT200808
WAT200809
WAT200839
WAT200510
WAT200511
WAT200826
Ion Chromatography
IC
WAT200814
WAT200815
WAT200845
WAT200816
WAT200817
WAT200843
* GHP GF and Nylon GF are Glass Fiber prefilters in combination with GHP and nylon filters.
** GF Glass Fiber only 1 m
84
Filtration
www.waters.com
SIMPLY CLEAR
SENSITIVIT Y IN ITS
PUREST FORM
86
87
CSH C18
USP
Class No.
pH Range
(Room Temp.)
Temperature
Limits
Particle
Size
Pore
Size
Surface
Area
Carbon
Load
L1
1-11
Low pH = 80 C
High pH = 45 C
1.7 m
130
185 m2/g
15%
Selectivity Features: General purpose reversed-phase column that offers excellent pH stability and rapid mobile-phase
re-equilibration for method development. Charged Surface Hybrid (CSH) technology enables superior peak shape and increased
loading capacity for basic compounds.
Bonding: Trifunctional C18 ligand, fully end-capped, bonded to a CSH particle substrate.
O Si
O
L11
CSH
Phenyl-Hexyl
L43
O
O Si
O
O
OO Si
OO Si
O
1-8
1-12
L7
1-12
CH 3
O Si
O
Polar Group
Si 3
OOCH
OO Si
O
O
Si 3
O CH
OSi
OO
L1
Polar Group
O CH3
OO Si
OO Si
O
O
O Si
O
O Si
OCH 3
OOSi
Polar Group
Si
O CH
3
O
2-11
BEH Phenyl
O
O Si
O
O
O Si
O
BEH HILIC
O
O Si
O
Low pH = 60 C
High pH = 45 C
1.7 m
130
185 m2/g
10%
Low pH = 80 C
High pH = 60 C
1.7 m
130
185 m2/g
18%
Low pH = 60 C
High pH = 60 C
1.7 m
130
185 m2/g
13%
Polar Group
Low pH = 50 C
High pH = 45 C
1.7 m
130
185 m2/g
17%
Selectivity Features: Alternate selectivity as compared to straight chain C18, particularly with phenolic analytes.
Compatible with 100% aqueous-phase composition.
Bonding: Monofunctional embedded polar C18, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
L11
1-12
O
O Si
O
O
CH 3
O Si
O Si
O
CH3
14%
Selectivity Features: General purpose column ideally suited for method development due to extreme pH stability and
applicability to the broadest range of compounds classes.
Bonding: Trifunctional C8, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
CH
O Si 3
OOSi
185 m2/g
Polar Group
Si 3
O CH
O
O Si
O
130
Selectivity Features: General purpose column ideally suited for method development due to extreme pH stability and
applicability to the broadest range of compound classes.
Bonding: Trifunctional C18, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
O Si
O
BEH C8
1.7 m
Selectivity Features: General purpose column that provides a very high degree of analyte selectivity, especially when using
low-pH mobile phases. Charged Surface Hybrid (CSH) technology enables superior peak shape and increased loading capacity
for basic compounds.
Bonding: Trifunctional propyl fluorophenyl ligand, non-endcapped, bonded to a CSH particle substrate.
L1
BEH C18
Low pH = 80 C
High pH = 45 C
Selectivity Features: General purpose alternative selectivity ligand that provides pi-pi interactions with polyaromatic compounds,
while maintaining excellent reproducibility at pH extremes. Charged Surface Hybrid (CSH) technology enables superior peak shape
and increased loading capacity for basic compounds.
Bonding: Trifunctional C6 Phenyl ligand, fully end-capped, bonded to a CSH particle substrate.
O
O Si
O
CSH
Fluoro-Phenyl
1-11
Low pH = 80 C
High pH = 60 C
1.7 m
130
185 m2/g
15%
Selectivity Features: Excellent method development column for alternate selectivity, particularly in regard to
polyaromatic compounds. Unique level of pH stability for a Phenyl bonded phase.
Bonding: Trifunctional C6 Phenyl, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
L3
1-9
Low pH = 45 C
High pH = 45 C
1.7 m
130
185 m2/g
unbonded
Selectivity Features: Excellent for retention of very polar, basic, water soluble analytes. Specifically designed and tested for
HILIC separations using mobile phases containing high concentrations of organic solvent.
Bonding: Unbonded Ethylene Bridged Hybrid (BEH) substrate.
O Si
O
88
www.waters.com
BEH Amide
O Si
O
Linker
NH 2
(Peptide Separation
Technology)
(Protein Separation
Technology)
O
CH
O Si 3
OOSi
CH 3
O CH
Si 3
Polar Group
CH3
2-11
Low pH = 90 C
High pH = 90 C
1.7 m
130
185 m2/g
12%
Selectivity Features: Rugged HILIC stationary phase designed to separate a wide range of very polar compounds. Especially good
at separating carbohydrates (saccharides) using high concentrations of organic modifier, elevated temperature and high pH. Compatible
with all modern detectors including MS, ELSD, UV and Fluorescence.
Bonding: Trifuncional amide bonded to Ethylene Bridged Hybrid (BEH) substrate.
L1
OO Si
O Si
O
O CH
Si
O
O Si
O
O CH
Si
O
O
Low pH = 80 C
High pH = 60 C
1.7 m
130
185 m2/g
18%
Low pH = 80 C
High pH = 60 C
1.7 m
300
90 m2/g
12%
1-10
Low pH = 80 C
High pH = 50 C
1.7 m
300
90 m2/g
8%
1-12
Low pH = 80 C
High pH = 60 C
1.7 m
130
185 m2/g
18%
Selectivity Features: pH and temperature stable, small pore, C18, UPLC column for synthetic DNA and RNA. Specifically QC tested
with synthetic DNA ladder.
Bonding: Trifunctional C18, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
O Si
O
2-11
3
Polar Group
OCH 3
O
O
Si
O Si
O Si
OCH
O 3
Linker
NH 2
Polar Group
O
O
O Si
O Si
OO
O Si
O
O
OH
OH
O
Si 3
O CH
O
OOSi
O Si
CH3
O
Low pH = 90 C
High pH = 90 C
1.7 m
130
185 m2/g
12%
Selectivity Features: A HILIC UPLC column for the LC and LC/MS analysis of fluorescently labeled glycans. Specifically QC
tested with 2-AB labeled human mAb IgG derived glycans.
Bonding: Trifunctional amide bonded to Ethylene Bridged Hybrid (BEH) substrate.
L33
HSS C18
1-12
OOSi
O
OO Si
OO Si
BEH200 SEC
1-12
C4 H 9
CH3
(Glycan Separation
Technology)
Carbon
Load
Selectivity Features: pH and temperature stable, wide pore, C4, UPLC column for proteins. Specifically QC tested with protein mixture.
Bonding: Proprietary monofunctional C4 bonding to Ethylene Bridged Hybrid (BEH) substrate.
O Si
CH 3 C4 H 9
BEH Glycan
Surface
Area
L26
OO Si
OO Si
O
CH 3
(Oligonucleotide
Separation Technology)
Pore
Size
Selectivity Features: pH and temperature stable, wide pore, C18, UPLC column for peptides. Specifically QC tested with peptide map.
Bonding: Trifunctional C18, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
Polar Group
CH3
O Si
BEH C18
Particle
Size
L1
O Si
BEH300 C4
Temperature
Limits
Selectivity Features: pH and temperature stable, small pore, C18, UPLC column for peptides. Specifically QC tested with peptide map.
Bonding: Trifunctional C18, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
O Si
O
BEH300 C18
pH Range
(Room Temp.)
L1
BEH130 C18
(Peptide Separation
Technology)
USP
Class No.
1-8
Low pH = 60 C
High pH = 60 C
1.7 m
200
216 m2/g
12%
Selectivity Features: A UPLC SEC column for proteins from 10,000 to 450,000 daltons. Specifically QC tested with protein standards.
Bonding: Diol bonded to a high pore volume Ethylene Bridged Hybrid (BEH) substrate.
Polar Group
L1
O
O
O
Si
O
O Si
O Si
O
O
O
1-8
Low pH = 45 C
High pH = 45 C
1.8 m
100
230 m2/g
15%
Selectivity Features: Ultra performance C18 chemistry, increased retention, superior peak shape, resists acid hydrolysis at low pH.
Designed for UPLC separations where silica-based C18 selectivities are desired.
Bonding: High coverage trifunctional C18, fully endcapped, bonded to High Strength Silica (HSS) UPLC particle substrate.
CH 3
O Si
CH3
HSS C18 SB
O
O
Si
O Si
O
O
Polar Group
L1
www.waters.com
O
O Si
O
Low pH = 45 C
High pH = 45 C
1.8 m
100
230 m2/g
8%
Selectivity Features: Unique, non-endcapped C18 chemistry designed specifically for method development scientists.
Offers unique Selectivity for Bases (SB) when operating under low pH conditions.
Bonding: Intermediate coverage tri-functionally bonded C18, no endcapping, bonded to High Strength Silica (HSS) UPLC particle substrate.
L1
HSS T3
2-8
2-8
Low pH = 45 C
High pH = 45 C
1.8 m
100
230 m2/g
11%
Selectivity Features: Aqueous mobile-phase compatible UPLC column designed for extreme retention.
Combines polar compound retention with UPLC efficiencies and performance.
Bonding: T3 (C18) bonding and endcapping, bonded to High Strength Silica (HSS) UPLC particle substrate.
89
90
www.waters.com
Particle Type
Particle Size
1.7 m
1.7 m
1.8 m
185 m2/g
185 m2/g
216 m2/g
90 m2/g
230 m2/g
C18, Phenyl-Hexyl,
Fluoro-Phenyl
C18, C8,
Shield RP18, Phenyl,
HILIC, Amide
Diol
C18, C4
Diol (1-8)
C18 (1-12);
C4 (1-10)
C18 (1-8);
C18 SB, T3 (2-8)
Surface Area
Available Chemistries
pH Range
0.10
0.08
2.0 %
Imipramine
AU
0.06
0.04
Amitriptyline
1.0 %
0.5 %
0.02
0.1 %
0.00
0.00
0.20
0.40
0.60
0.80
1.00
1.20
1.40
0.10
1.60
1.80
2.00 min
Core-shell
Technology
0.08
Imipramine
AU
0.04
2.0 %
1.0 %
0.5 %
0.02
0.1 %
0.00
0.00
0.20
0.40
0.60
0.80
1.00
1.20
1.40
1.60
1.80
2.00 min
The increased and improved loadability possible with CSH Technology permits the separation, identification and quantitation of closely
eluting impurities or degradants. In this example, a high concentration (0.5 mg/mL) of imipramine was repeatedly injected while the
concentration of amitriptyline was increased with each injection.
www.waters.com
91
Temperature:
Injection Volume:
Detection:
Flow Rate:
System:
Compounds:
1. T hiourea
2. Resorcinol
3. Metoprolol
4. 3-Nitrophenol
5. 2-Chlorobenzoic acid
6. Amitriptyline
7. Diethylphthalate
8. Fenoprofen
9. Dipropylphthalate
10. Pyrenesulfonic acid
Compounds:
1.
2.
3.
4.
5.
6.
7.
8.
9.
Aminopyrazine
Pindolol
Quinine
Quinine impurity
Labetalol
Diltiazem impurity
Diltiazem
Verapamil
Amitriptyline
0.02
7
5 m
0.16
9
3
0.12
8
0.08
1
3.5 m
0.04
6
4
0.00
0.00
0.50
1.00
1.50
2.00
2.50
3.00 min
1.7 m
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0 min
92
www.waters.com
Compounds
1. HMX
2. RDX
3. 1,3,5-TNB
4. 1,3-DNB
5. NB
6. Tetryl
7. TNT
8. 2-Am-4,6 DNT
9. 4-Am-2,6 DNT
10. 2,4 DNT
11. 2,6-DNT
12. 2-NT
13. 4-NT
14. 3-NT
72% A; 28% B
5 L
10 g/mL
50 C
UV @ 254 nm
ACQUITY UPLC with PDA detector
3
1
2
5
10
8
11
10
8
6 9
11
BEH C8
12 13 14
3
1
8
9
13
10
6
11
10
11
12 13
14
14
12
4
2
BEH C18
12 13 14
1 2
BEH Phenyl
7
10
12
14
min
Injection Volume:
Sample Conc.:
Sample Diluent:
Temperature:
Detection:
Instrument:
2. Morphine 3-fl-D-Glucronide
C H3
N
C H3
OH
O O
HO
OH
HO
OH
UV @ 280 nm
ACQUITY UPLC with TUV Detector
V0
0.0
www.waters.com
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0 min
93
Carbohydrate Analysis
BEH particle tec hnology, in combination with a trifunctionallybonded amide phase, provides exceptional column lifetime, thus
improving assay robustness.
Injection Volume:
Sample Diluent:
Column Temp.:
Weak Needle Wash:
Detection:
Sampling Rate:
Filter Time Constant:
Instrument:
Column:
Part Number:
Mobile Phase A:
Mobile Phase B:
Flow Rate:
Gradient:
Injection Volume:
Sample Diluent:
Column Temp.:
Needle Wash:
Instrument:
25 g/mL
50 g/mL
50 g/mL
50 g/mL
50 g/mL
25 g/mL
50 g/mL
25 g/mL
Ionization Mode:
Capillary:
Cone Voltage:
Source Temp.:
Desolv. Temp.:
Desolv. Gas Flow:
SIR m/z:
Dwell Time:
Compounds:
1. Fructose
2. Glucose
3. Sucrose
4. Maltose
5. Maltotriose
6. Maltotetraose
7. Maltopentaose
8. Maltohexahose
9. Maltoheptaose
10 g/mL
10 g/mL
10 g/mL
10 g/mL
10 g/mL
10 g/mL
10 g/mL
10 g/mL
10 g/mL
2
7
7
3
min
94
7 min
www.waters.com
Injection Volume:
Sample Diluent:
Sample Conc.:
Temperature:
Detection:
Instrument:
HSS C18
Compounds
1. Caftaric acid
2. Chlorogenic acid
3. Cynarin
4. Echinacoside
5. Cichoric acid
5
4
HSS C18 SB
HSS T3
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0 min
By combining ultra-efficient particle tec hnology, low dispersion instrumentation and different column ligands, chromatographers can efficiently
and effectively develop robust methods faster than ever before.
Selectivity
ln (amintriptyline/toluene) pH 3.0
HSS C 18 SB
-0.2
BEH300 C4
BEH Phenyl
-0.4
-0.6
BEH C8
-0.8
BEH C18
BEH300 C18
-1.0
CSH Phenyl-Hexyl
-1.2
CSH Fluoro-Phenyl
-1.4
BEH
Shield RP18
HSS T3
HSS C 18
CSH C18
-1.6
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
Hydrophobicity ln k (toluene)
35% MeCN/65% 15.4 mM HCOONH4, pH 3.0
www.waters.com
95
FEATURE
BENEFIT
Column:
Part Number:
Pre-column:
Part Number:
Mobile Phase A:
Mobile Phase B:
Flow Rate:
Gradient:
Injection Volume:
Temperature:
Detection:
Sampling Rate:
Time Constant:
Instrument:
ACQUITY SQ Detector
ES+
Capillary:
Cone:
Source Temp:
Desolvation Temp:
Cone Gas Flow:
Desolvation Gas Flow:
SIR:
MS Interscan Delay:
Dwell:
3.5 kV
35 V
150 C
500 C
50 L/Hr
850 L/Hr
373.2 m/z, 415.2 m/z
0.005 sec
0.005 sec
OCH3
N
N
0.8
1.0
Diltiazem
SIR m/z 415.2
Without VanGuard pre-column
With VanGuard pre-column
0.6
96
OH
1.2
1.4
1.6
1.8
O
O O
2.0 min
www.waters.com
3.2
4.0
4.8
5.6
min
1.7 m
1.8 m
1.7 m
12
3.5, 5.0 m
3.5, 5.0 m
20
24 min
16
12
16
20
24 min
A separation of clozapine and its related compounds is used to demonstrate the exceptional scalability between
different size chromatographic particles and instrument platforms to facilitate a successful method transfer.
Retention Time Relative to Clozapine
www.waters.com
Separation Mode
Instrument
Impurity D
Impurity C
Impurity A
Impurity B
UPLC
0.867
0.890
0.939
1.500
HPLC
0.865
0.895
0.950
1.513
HPLC
0.867
0.898
0.951
1.507
97
* Eac h kit contains one UPLC column and one HPLC column. T he ACQUIT Y UPLC
columns calculator can be downloaded from the ACQUITY UPLC online community
at: www.waters.com/myuplc
Package Name
Chemistries
The widest selectivity offering for method development at low and high pH.
Best choice for low ionic strength additives (i.e., formic acid)
C18 columns that differ in silanol activity and hydrophobicity within the US
Pharmacopeia L1 classification
Straight-chain alkyl C18 columns that differ in silanol activity, shape, selectivity and
hydrophobicity, and exhibit no MS bleed
Offers the widest separation selectivity by combining CSH reversed-phase and HILIC
stationary phases to retain analytes encompassing a broad range of polarity
Effortlessly develop HILIC methods at low pH (bases) or high pH (acids) for polar
and/or ionizable compounds
98
www.waters.com
Chemistry
Particle
Size
Dimensions
Part No.
1 pack
Part No.
3 pack
CSH C18
CSH C18
CSH C18
CSH C18
CSH C18
CSH C18
CSH C18
CSH C18
CSH C18
CSH C18
CSH C18
CSH C18
CSH C18
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Fluoro-Phenyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
CSH Phenyl-Hexyl
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
186005292
186005293
186005294
186005295
186005296
186005620
186005297
186005298
186005299
186005300
186005623
186005301
186005302
186005349
186005347
186005348
186005350
186005351
186005622
186005352
186005353
186005354
186005355
186005625
186005356
186005357
186005404
186005402
186005403
186005405
186005406
186005621
186005407
186005408
186005409
186005410
186005624
186005411
186005412
176002136
176002137
176002138
176002139
176002140
176002141
176002142
176002143
176002144
176002145
176002146
176002150
176002148
176002149
176002151
176002152
176002153
176002154
176002155
176002156
176002157
176002158
176002161
176002159
176002160
176002162
176002163
176002164
176002165
176002166
176002167
176002168
176002169
Chemistry
Particle
Size
Dimensions
Part No.
1 pack
Part No.
3 pack
BEH C18
BEH C18
BEH C18
BEH C18
BEH C18
NEW BEH C18
BEH C18
BEH C18
BEH C18
BEH C18
NEW BEH C18
BEH C18
BEH C18
BEH Shield RP18
BEH Shield RP18
BEH Shield RP18
BEH Shield RP18
BEH Shield RP18
NEW BEH Shield RP18
BEH Shield RP18
BEH Shield RP18
BEH Shield RP18
BEH Shield RP18
NEW BEH Shield RP18
BEH Shield RP18
BEH Shield RP18
BEH C8
BEH C8
BEH C8
BEH C8
BEH C8
NEW BEH C8
BEH C8
BEH C8
BEH C8
BEH C8
NEW BEH C8
BEH C8
BEH C8
BEH Phenyl
BEH Phenyl
BEH Phenyl
BEH Phenyl
BEH Phenyl
NEW BEH Phenyl
BEH Phenyl
BEH Phenyl
BEH Phenyl
BEH Phenyl
NEW BEH Phenyl
BEH Phenyl
BEH Phenyl
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
186002344
186002346
186002347
186002349
186002350
186005604
186002352
186002353
186004659
186004660
186005609
186004661
186004690
186002851
186002852
186003373
186003909
186002853
186005605
186002854
186003376
186004667
186004668
186005610
186004669
186004670
186002875
186002876
186003374
186003910
186002877
186005606
186002878
186003377
186004663
186004664
186005611
186004665
186004666
186002882
186002883
186003375
186003911
186002884
186005607
186002885
186003378
186004671
186004672
186005612
186004673
186004674
176000861
176000862
176001044
176001304
176000863
176000864
176001048
176001794
176001795
176001796
176001797
176000874
176000875
176001045
176001309
176000876
176000877
176001049
176001802
176001803
176001804
176001805
176000882
176000883
176001046
176001310
176000884
176000885
176001050
176001798
176001799
176001800
176001801
176000905
176000906
176001047
176001311
176000907
176000908
176001051
176001806
176001807
176001808
176001809
www.waters.com/amidemethod
T his interactive tool requires that the user has Adobe Flash Player 8.
www.waters.com
99
Chemistry
Particle
Size
Dimensions
Part No.
1 pack
Part No.
3 pack
BEH HILIC
BEH HILIC
BEH HILIC
BEH HILIC
NEW BEH HILIC
BEH HILIC
BEH HILIC
BEH HILIC
NEW BEH HILIC
BEH HILIC
BEH HILIC
BEH Amide
BEH Amide
BEH Amide
BEH Amide
BEH Amide
NEW BEH Amide
BEH Amide
BEH Amide
BEH Amide
BEH Amide
NEW BEH Amide
BEH Amide
BEH Amide
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
186003457
186003458
186003459
186003460
186005608
186003461
186003462
186004675
186005613
186004676
186004677
186004848
186004849
186004850
186004839
186004800
186005657
186004801
186004802
186004803
186004804
186005658
186004805
186004806
176001089
176001090
176001091
176001092
176001093
176001094
176001810
176001811
176001812
176001914
176001915
176001916
176001906
176001907
176001908
176001909
176001910
176001911
176001912
176001913
Chemistry
Particle
Size
Dimensions
Part No.
1 pack
Part No.
3 pack
HSS T3
HSS T3
HSS T3
HSS T3
HSS T3
NEW HSS T3
HSS T3
HSS T3
HSS T3
HSS T3
NEW HSS T3
HSS T3
HSS T3
HSS C18
HSS C18
HSS C18
HSS C18
HSS C18
NEW HSS C18
HSS C18
HSS C18
HSS C18
HSS C18
NEW HSS C18
HSS C18
HSS C18
HSS C18 SB
HSS C18 SB
HSS C18 SB
HSS C18 SB
HSS C18 SB
NEW HSS C18 SB
HSS C18 SB
HSS C18 SB
HSS C18 SB
HSS C18 SB
NEW HSS C18 SB
HSS C18 SB
HSS C18 SB
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
186003535
186003536
186003537
186003944
186003538
186005614
186003539
186003540
186004678
186004679
186005617
186004680
186004681
186003529
186003530
186003531
186003987
186003532
186005615
186003533
186003534
186004682
186004683
186005618
186004684
186004685
186004114
186004115
186004116
186004117
186004118
186005616
186004119
186004120
186004686
186004687
186005619
186004826
186004689
176001127
176001129
176001130
176001375
176001131
176001132
176001133
176001813
176001814
176001815
176001816
176001121
176001122
176001123
176001398
176001124
176001125
176001126
176001817
176001818
176001819
176001820
176001556
176001557
176001558
176001559
176001560
176001561
176001562
176001821
176001822
176001823
176001824
100
Description
Beginners Guide to UPLC
Quest for Ultra Performance in Liquid Chromatography
NEW Comprehensive Guide to HILIC
Part No.
715002099
715002098
715002531
www.waters.com
www.waters.com
Qty/Pk
Chemistries
Particle Size(s)
Dimensions
Part No.
4/pk
2.1 x 50 mm
176002123
4/pk
2.1 x 100 mm
176002124
4/pk
3.0 x 50 mm
176002125
4/pk
3.0 x 100 mm
176002126
4/pk
BEH: 1.7 m
2.1 x 50 mm
176001042
4/pk
BEH: 1.7 m
2.1 x 100 mm
176001043
4/pk
BEH: 1.7 m
3.0 x 50 mm
176001881
4/pk
BEH: 1.7 m
3.0 x 100 mm
176001882
4/pk
4/pk
4/pk
4/pk
4/pk
4/pk
4/pk
4/pk
4/pk
4/pk
4/pk
4/pk
4/pk
4/pk
4/pk
4/pk
2.1 x 50 mm
2.1 x 100 mm
3.0 x 50 mm
3.0 x 100 mm
2.1 x 50 mm
2.1 x 100 mm
3.0 x 50 mm
3.0 x 100 mm
2.1 x 50 mm
2.1 x 100 mm
3.0 x 50 mm
3.0 x 100 mm
2.1 x 50 mm
2.1 x 100 mm
3.0 x 50 mm
3.0 x 100 mm
176001603
176001604
176001883
176001884
176001605
176001606
176001885
176001886
176001607
176001608
176001887
176001888
176001609
176001610
176001889
176001890
4/pk
2.1 x 50 mm
176002127
4/pk
2.1 x 100 mm
176002128
4/pk
3.0 x 50 mm
176002129
4/pk
3.0 x 100 mm
176002130
4/pk
4/pk
4/pk
4/pk
2/pk
2/pk
2/pk
2/pk
2.1 x 50 mm
2.1 x 100 mm
3.0 x 50 mm
3.0 x 100 mm
2.1 x 50 mm
2.1 x 100 mm
3.0 x 50 mm
3.0 x 100 mm
176001959
176001960
176001961
176001962
176001963
176001964
176001965
176001966
101
HPLC Column
4.6 mm I.D.
Part No.
50 mm, 1.7 m
50 mm, 1.7 m
150 mm, 5 m
150 mm, 5 m
186005529
186005530
50 mm, 1.7 m
150 mm, 5 m
186005531
50 mm, 1.7 m
50 mm, 1.7 m
50 mm, 1.7 m
50 mm, 1.8 m
50 mm, 1.8 m
50 mm, 1.8 m
50 mm, 1.7 m
150 mm, 5 m
150 mm, 5 m
150 mm, 5 m
150 mm, 5 m
150 mm, 5 m
150 mm, 5 m
100 mm, 3.5 m
186004958
186004959
186004960
186004961
186004962
186004963
186005532
50 mm, 1.7 m
186005533
50 mm, 1.7 m
186005534
50 mm, 1.7 m
50 mm, 1.7 m
50 mm, 1.7 m
50 mm, 1.7 m
50 mm, 1.8 m
50 mm, 1.8 m
50 mm, 1.8 m
186004964
186004965
186004966
186004967
186004968
186004969
186004970
186005535
186005536
186005537
186004971
186004972
186004973
186004974
186004975
186004976
186004977
Package Name
NEW CSH C18 1.7 to 5 m Method Transfer kit
NEW CSH Phenyl-Hexyl 1.7 to 5 m Method Transfer kit
NEW CSH Fluoro-Phenyl 1.7 to 5 m
Method Transfer kit
BEH C18 1.7 to 5 m Method Transfer kit
BEH Shield RP18 1.7 to 5 m Method Transfer kit
BEH HILIC 1.7 to 5 m Method Transfer kit
HSS C18 1.8 to 5 m Method Transfer kit
HSS T3 1.8 to 5 m Method Transfer kit
HSS C18 SB 1.8 to 5 m Method Transfer kit
NEW CSH C18 1.7 to 3.5 m Method Transfer kit
NEW CSH Phenyl-Hexyl 1.7 to 3.5 m
Method Transfer kit
NEW CSH Fluoro-Phenyl 1.7 to 3.5 m
Method Transfer kit
BEH C18 1.7 to 3.5 m Method Transfer kit
BEH Shield RP18 1.7 to 3.5 m Method Transfer kit
BEH HILIC 1.7 to 3.5 m Method Transfer kit
BEH Amide 1.7 to 3.5 m Method Transfer kit
HSS C18 1.8 to 3.5 m Method Transfer kit
HSS T3 1.8 to 3.5 m Method Transfer kit
HSS C18 SB 1.8 to 3.5 m Method Transfer kit
NEW CSH C18 1.7 to 3.5 m
High Rs Method Transfer kit
NEW CSH Phenyl-Hexyl 1.7 to 3.5 m
High Rs Method Transfer kit
NEW CSH Fluoro-Phenyl 1.7 to 3.5 m
High Rs Method Transfer kit
BEH C18 1.7 to 3.5 m High Rs Method Transfer kit
BEH Shield RP18 1.7 to 3.5 m High
Rs Method Transfer kit
BEH HILIC 1.7 to 3.5 m High Rs Method Transfer kit
BEH Amide 1.7 to 3.5 m High Rs Method Transfer kit
HSS C18 1.8 to 3.5 m High Rs Method Transfer kit
HSS T3 1.8 to 3.5 m High Rs Method Transfer kit
HSS C18 SB 1.8 to 3.5 m High Rs Method Transfer kit
Chemistry
NEW CSH C18
NEW CSH C18
NEW CSH
Phenyl-Hexyl
NEW CSH
Phenyl-Hexyl
NEW CSH
Fluoro-Phenyl
NEW CSH
Fluoro-Phenyl
BEH C18
BEH C18
BEH C8
BEH C8
BEH Shield RP18
BEH Shield RP18
BEH Phenyl
BEH Phenyl
BEH HILIC
BEH HILIC
BEH Amide
BEH Amide
HSS T3
HSS T3
HSS C18
HSS C18
HSS C18 SB
HSS C18 SB
Particle
Size
Column
Length
Part No.
2.1 mm I.D.
Part No.
3.0 mm I.D.
1.7 m
1.7 m
50 mm
100 mm
186005571
186005572
186005573
186005574
1.7 m
50 mm
186005579
186005581
1.7 m
100 mm
186005580
186005582
1.7 m
50 mm
186005575
186005577
1.7 m
100 mm
186005576
186005578
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
1.8 m
50 mm
100 mm
50 mm
100 mm
50 mm
100 mm
50 mm
100 mm
50 mm
100 mm
50 mm
100 mm
50 mm
100 mm
50 mm
100 mm
50 mm
100 mm
186004044
186004045
186004046
186004047
186004048
186004049
186004050
186004052
186004053
186004054
186004807
186004808
186004055
186004056
186004057
186004058
186004137
186004138
186004691
186004692
186004693
186004694
186004695
186004696
186004697
186004698
186004699
186004700
186004809
186004810
186004701
186004702
186004703
186004704
186004705
186004709
* Each kit contains one UPLC column and one HPLC column. The ACQUITY UPLC Columns
Calculator can be downloaded from the ACQUITY UPLC Online Community at www.waters.com/myuplc
Chemistry
Particle
Size
Dimensions
Part No.
3 pack
1.7 m
1.7 m
1.7 m
2.1 x 5 mm
2.1 x 5 mm
2.1 x 5 mm
186005303
186005358
186005413
Description
BEH C18
BEH Shield RP18
BEH C8
BEH Phenyl
BEH HILIC
BEH Amide
HSS C18
HSS C18 SB
HSS T3
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.8 m
1.8 m
1.8 m
2.1 x 5 mm
2.1 x 5 mm
2.1 x 5 mm
2.1 x 5 mm
2.1 x 5 mm
2.1 x 5 mm
2.1 x 5 mm
2.1 x 5 mm
2.1 x 5 mm
186003975
186003977
186003978
186003979
186003980
186004799
186003981
186004136
186003976
102
Part No.
205000343
700002775
www.waters.com
Amber
Max Recovery
Amber Max
300 L PP
750 L PP
Clear Glass
with Septumless Cap
600000668CV
600000669CV
600000670CV
600000755CV
186003886C
186000307C
186000847C
186000327C
186000307DV
186000847DV
186000327DV
186004132C
186004133C
186004168C
186004132C
Combination Packs
Bonded Pre-slit Silicone/PTFE Septum
186002639
186004112
186005221
1600 L
1600 L
1100 L
1100 L
210 L
530 L
1600 L
Residual
165 L
165 L
22 L
22 L
20 L
70 L
165 L
www.waters.com
Clear 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with Quick Thread Design,
(6 mm Opening, 9 mm Cap).
Amber 12 x 32, Type 1, 51-Expansion Glass Screw Neck with Quick Thread Design,
(6 mm Opening, 9 mm Cap).
Waters Maximum Recovery Vial, 12 x 32, Type 1, 33-Expansion Glass, Screw Neck
with Quick Thread Design (6 mm Opening, 9 mm Cap).
Clear 12 x 32, Type 1, 33-Expansion Glass, Screw Neck with Quick Thread Design,
(6 mm Opening, 9 mm Septumless Cap).
Part No.
405003743
103
384-Well Plates
186002643
186002481
186002482
186002632
100
50
50
50
50
350 L
800 L
2 mL
250 L
100 L
186002483
186002483
186002484
186002788
186002788
186002788
186002788
186002788
186002789
186002789
186002789
186002789
186002789
35 L
15 L
20 L
15 L
15 L
Part No.
Max Volume
Residual Volume
15 L
Pack Size
Well Volume
186002631
Sealing Options
1860014381
186001437 (DV)2
650 L
186001436 (DV)2
850 L
15 L
186000349 (DV)2
650 L
15 L
186000855 (DV)2
850 L
15 L
Sealing Cap for 700 L Glass Insert Square Well Seals Against the Well Wall PTFE/Silicone, 5/pk
186000857
Sealing Cap for 1 mL Glass Inserts Seals in the Glass PTFE/Silicone, 10/pk
186000856
When (DV) appears beside the part number, a deactivated version of this product can be ordered by adding DV to the right of the part number.
Waters ACQUITY UPLC System Sample Manager Version 1.30 and later
Sample Needle
Vial Depth
Typical 2 mL Vial
2 mm Default Offset
104
www.waters.com
Heat Sealer
Part No.
115 Volt
186002786
240 Volt
186002787
Solvent Range
Clear Polyester
Heat Seal
From -80 C to 80 C
From -200 C to 90 C
Recommendation
T he aluminum foil laminate heat seal is good for most solvents used
in laboratories. For applications requiring DMSO, the plates should be
stored at 4 C.
Position the seal with the white side facing up. Apply heat using the Waters
Heat Sealer for 2 to 3 seconds in both directions following the instructions
found on page 9 in the heater manual, part number: 720001330EN.
Both of these seals can be peeled off by hand. For plate storage, apply a
new unpierced seal or polypropylene cap mat.
www.waters.com/vialsselector
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Recommended Supplies for Use in Conjunction With the ACQUITY UPLC System
Filters
Waters recommends you filter buffers and samples using a 0.2 m filter
prior to use. Filtration protects the column and instrument components
from a build-up of particulate matter, improving column lifetime and
minimizing system down time. 0.2 m GHP Filters are recommended for
filtering aqueous, non-polar solvents and proteins.
Ostro sample preparation products provide a novel solution for the cleanup
of phospholipids in plasma and serum. Requiring minimal to no method
development and using a simple protocol, this technology can be quickly
implemented in order to optimize your laboratorys workflow. Providing
cleaner, more reproducible extracts than competitive phospholipid removal
devices or techniques, the Ostro 96-well plate allows for more sensitive
analyses, increased sample throughput, and reduced instrument downtime.
Description
Part No.
186003524
WAT097962
WAT097964
www.waters.com
Part No.
NEW Ostro Sample Preparation 96-Well Plate (25 mg), single pack
186005518
186003873
186002448
105
[ Analytical Chromatography ]
Introduction ...........................................................................................................................................................................................108
Analytical Column Listing .......................................................................................................................................................................109
USP L Column Listing ...........................................................................................................................................................112
Physical Characteristics of HPLC
and UPLC Packing Materials ....................................................................................................................................................116
HPLC Columns Theory, Technology, and Practice ...................................................................................................................117
Product and Ordering Information
NEW XSelect Columns ..............................................................................................................................................................118
XBridge Columns ......................................................................................................................................................................122
SunFire Columns .......................................................................................................................................................................129
Atlantis Columns......................................................................................................................................................................132
NEW High Strength Silica (HSS) HPLC Columns .....................................................................................................................138
XTerra Columns ........................................................................................................................................................................140
Symmetry Columns ..................................................................................................................................................................144
Waters Spherisorb Columns .....................................................................................................................................................152
Nova-Pak Packings ....................................................................................................................................................................155
Bondapak/Bondapak Packings................................................................................................................................................158
Delta-Pak Columns ...................................................................................................................................................................159
Porasil/Porasil Silica Packings ...............................................................................................................................................160
Resolve Columns ......................................................................................................................................................................161
Shodex RSpak Polymer Reversed-Phase Columns...................................................................................................................162
Guard Products ..........................................................................................................................................................................163
Waters Cartridge and Guard Column Guide ............................................................................................................................164
Paired-Ion Chromatography (PIC Reagents) .............................................................................................................................166
Applications Specific
Sugar and Carbohydrate Analysis ...........................................................................................................................................167
Fatty Acid Analysis ..................................................................................................................................................................171
Pesticide Analysis ....................................................................................................................................................................171
Waters PAH Columns ...............................................................................................................................................................172
Triglycerides and Cholesterol Analysis ..................................................................................................................................173
Fermentation Analysis, Organic Acids,
Alcohols, and Carbohydrates ...................................................................................................................................................173
Ion Analysis ..............................................................................................................................................................................174
Transition Metal Analysis........................................................................................................................................................175
Lanthanide Analysis .................................................................................................................................................................175
Gas-Chromatography Packings.................................................................................................................................................................176
106
[ Preparative Chromatography ]
Column PerformanceIdentifying the Problem ....................................................................................................................................177
Waters OBD Column Design ....................................................................................................................................................................177
Column Stability and ReliabilityLong, Predictable Lifetimes ...........................................................................................................178
ScalabilityAnalytical Performance in a Prep OBD Column ..............................................................................................................178
Selecting the Column Packing .................................................................................................................................................................179
Waters AutoPurification System .............................................................................................................................................................179
Choosing Preparative Columns ................................................................................................................................................................180
Product and Ordering Information
NEW XSelect OBD Columns......................................................................................................................................................181
XBridge Prep OBD Columns .....................................................................................................................................................183
SunFire Prep OBD Columns ......................................................................................................................................................185
Atlantis Prep OBD Columns .....................................................................................................................................................187
XTerra Prep OBD Columns........................................................................................................................................................189
Symmetry Columns ..................................................................................................................................................................190
SymmetryShield Columns ........................................................................................................................................................190
Symmetry300 Columns ..........................................................................................................................................................190
SymmetryPrep Columns ...........................................................................................................................................................190
Waters Spherisorb Columns .....................................................................................................................................................191
Prep Nova-Pak HR Columns......................................................................................................................................................191
Bondapak/Porasil Columns ......................................................................................................................................................191
Delta-Pak Columns ...................................................................................................................................................................192
Preparative Bulk Material.........................................................................................................................................................192
Radial Compression Technology
Radial Compression Modules ...................................................................................................................................................193
Radial Compression Column Segments ..................................................................................................................................193
PrepPak Cartridges ....................................................................................................................................................................193
Accessories and Spare Parts ....................................................................................................................................................195
107
aNaLYTICaL ChROMaTOGRaPhY
Introduction
Featuring
CSH Technology
Choosing an HPLC column can be a very difficult process. T he Chromatographic Packing Material (particle brand name and functional group)
plays a major role in the success of a separation, both initially, when the
method is first developed, as well as long term, if the method will need
to be reproduced for many years.
Featuring
BEH Technology
108
Analytical Chromatography
Description
Beginners Guide to Liquid Chromatography
NEW Comprehensive Guide to HILIC
Part No.
715001531
715002531
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
USP
Class No.
pH Range
(Room Temp.)
Temperature
Limits
Particle
Size
Pore
Size
Surface
Area
Carbon
Load
L1
1-11
Low pH = 80 C
High pH = 45 C
3.5, 5 m
130
185 m2/g
15%
CSH C18
Selectivity Features: General purpose reversed-phase column that offers excellent pH stability and rapid mobile-phase
re-equilibration for method development. Charged Surface Hybrid (CSH) technology enables superior peak shape and increased
loading capacity for basic compounds.
Bonding: Trifunctional C18 ligand, fully end-capped, bonded to a CSH particle substrate.
O Si
O
L11
CSH
Phenyl-Hexyl
O
O Si
O
Low pH = 80 C
High pH = 45 C
3.5, 5 m
130
185 m2/g
14%
Selectivity Features: General purpose alternative selectivity ligand that provides pi-pi interactions with polyaromatic compounds,
while maintianing excellent reproducibility at pH extremes. Charged Surface Hybrid (CSH) technology enables superior peak shape
and increased loading capacity for basic compounds.
Bonding: Trifunctional C6 Phenyl ligand, fully end-capped, bonded to a CSH particle substrate.
O
O Si
O
L43
CSH
Fluoro-Phenyl
1-11
1-8
Low pH = 60 C
High pH = 45 C
3.5, 5 m
130
185 m2/g
10%
Selectivity Features: General purpose column that provides a very high degree of analyte selectivity, especially when using
low-pH mobile phases. Charged Surface Hybrid (CSH) technology enables superior peak shape and increased loading capacity
for basic compounds.
Bonding: Trifunctional propyl fluorophenyl ligand, non-endcapped, bonded to a CSH particle substrate.
C18
1-12
L7
1-12
CH 3
O Si
O
CH
O Si 3
Polar Group
OOSi
18%
Low pH = 60 C
High pH = 60 C
2.5, 3.5,
5, 10 m
130
185 m2/g
13%
Polar Group
L1
2-11
O
O
O Si
OCH
O Si
Polar Group
O CH
3
Si
OO
O
OSi
O
O Si
O
O
O
O Si
CH 3
O
O Si
O
CH
OO Si 3
Polar Group
OOSi
O
Phenyl
185 m2/g
CH3
O Si
O
O
O Si
Shield RP18
130
Selectivity Features: General purpose column ideally suited for method development due to extreme pH stability and
applicability to the broadest range of compound classes.
Bonding: Trifunctional C8, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
O
O
O Si
O
O
Si
O CH
2.5, 3.5,
5, 10 m
Selectivity Features: General purpose column ideally suited for method development due to extreme pH stability and
applicability to the broadest range of compound classes.
Bonding: Trifunctional C18, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
O Si
O
O Si
O
O
O Si
O
C8
Low pH = 80 C
High pH = 60 C
O
Si
O CH
3
OOSi
CH3
O
O Si
O
Polar Group
Low pH = 50 C
High pH = 45 C
2.5, 3.5,
5, 10 m
130
185 m2/g
17%
Selectivity Features: Alternate selectivity as compared to straight chain C18, particularly with phenolic analytes.
Compatible with 100% aqueous-phase composition.
Bonding: Monofunctional embedded polar C18, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
L11
1-12
Low pH = 80 C
High pH = 60 C
2.5, 3.5, 5 m
130
185 m2/g
15%
Selectivity Features: Excellent method development column for alternate selectivity, particularly in regard to polyaromatic
compounds. Unique level of pH stability for a Phenyl bonded phase.
Bonding: Trifunctional C6 Phenyl, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
O
O Si
O
HILIC
O Si
O
L3
1-9
Low pH = 45 C
High pH = 45 C
2.5, 3.5,
5 m
130
185 m2/g
unbonded
Selectivity Features: Excellent for retention of very polar, basic, water soluble analytes. Specifically designed and tested for
HILIC separations using mobile phases containing high concentrations of organic solvent.
Bonding: Unbonded Ethylene Bridged Hybrid (BEH) substrate.
O Si
O
www.waters.com
Analytical Chromatography
109
aNaLYTICaL ChROMaTOGRaPhY
USP
Class No.
pH Range
(Room Temp.)
Temperature
Limits
Particle
Size
Pore
Size
Surface
Area
Carbon
Load
2-11
Low pH = 90 C
High pH = 90 C
3.5 m
130
185 m2/g
12%
Amide
O Si
O
Linker
NH 2
Selectivity Features: Rugged HILIC stationary phase designed to separate a wide range of very polar compounds. Especially good at
separating carbohydrates (saccharides) using high concentrations of organic modifier, elevated temperature and high pH. Compatible with all
modern detectors including MS, ELSD, UV and Fluorescence.
Bonding: Trifuncional amide bonded to Ethylene Bridged Hybrid (BEH) substrate.
L1
BEH130 C18
1-12
O
O Si
O
(Peptide Separation
Technology)
Low pH = 80 C
High pH = 60 C
3.5, 5, 10 m
130
185 m2/g
18%
Selectivity Features: pH and temperature stable, small pore, C18, UPLC column for peptides. Specifically QC tested with peptide map.
Bonding: Trifunctional C18, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
O
O Si
O
BEH300 C18
L1
OCH 3
Si
O Si
(Peptide Separation
Technology)
Polar Group
O
CH3
O
O
O Si
O SiCH
3
O
O
O Si
BEH300 C4
1-12
L26
C4 H 9
1-10
CH33
CH
OO SiSi
Polar
C4Group
H9
CH33
CH
(Oligonucleotide
Separation Technology)
300
90 m2/g
12%
Low pH = 80 C
High pH = 50 C
3.5 m
300
90 m2/g
8%
Selectivity Features: pH and temperature stable, wide pore, C4, UPLC column for proteins. Specifically QC tested with protein mixture.
Bonding: Proprietary monofunctional C4 bonding to Ethylene Bridged Hybrid (BEH) substrate.
O
O
O Si
O Si
O
O
L1
BEH C18
3.5, 5, 10 m
Selectivity Features: pH and temperature stable, wide pore, C18, UPLC column for peptides. Specifically QC tested with peptide map.
Bonding: Trifunctional C18, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
CH3
(Protein Separation
Technology)
Low pH = 80 C
High pH = 60 C
1-12
Low pH = 80 C
High pH = 60 C
2.5 m
130
185 m2/g
18%
Selectivity Features: pH and temperature stable, small pore, C18, UPLC column for synthetic DNA and RNA. Specifically QC tested
with synthetic DNA ladder.
Bonding: Trifunctional C18, fully endcapped, bonded to Ethylene Bridged Hybrid (BEH) substrate.
O
O Si
O
O
O
O Si
O Si
O
O
O Si
Polar Group
CH3
L1
O
O Si
O
C18
2-8
Low pH = 50 C
High pH = 40 C
2.5, 3.5,
5, 10 m
100
340 m2/g
16%
Selectivity Features: General purpose method development column. Very high loading capacity, particularly for basic analytes
in low pH mobile phases. Ideally suited for purification and impurity profile assays.
Bonding: Difunctional C18, fully endcapped, bonded to high purity silica substrate.
Si
O
O
Si
O
OO
O Si Si
OO
L7
CH 3
C8
O Si
O CH
Si
Polar Group
Low pH = 40 C
High pH = 40 C
2.5, 3.5,
5, 10 m
100
340 m2/g
12%
Selectivity Features: General purpose method development column. Very high loading capacity, particularly for basic analytes
in low pH mobile phases. Less hydrophobic, therefore, less retentive than C18 for most analytes.
Bonding: Difunctional C8, fully endcapped, bonded to high purity silica substrate.
O
O Si
CH 3
O
O Si
2-8
Polar Group
CH3
O
O Si
O
O
O Si
O
O
O Si
O
110
Analytical Chromatography
www.waters.com
ANALYTICAL CHROMATOGRAPHY
USP
Class No.
pH Range
(Room Temp.)
Temperature
Limits
Particle
Size
L1
2-8
Low pH = 45 C
High pH = 45 C
3, 5, 10 m
Pore
Size
Surface
Area
Carbon
Load
100
330 m2/g
14%
O Si
O
O
O Si
O
T3
O
O Si
OCH
O Si
Polar Group
CH3
O
Selectivity Features: Retention of polar compounds, compatible with 100% aqueous mobile phases, superior stability under
low pH conditions. Specifically designed for enhanced retention of polar analytes.
Bonding: T3 (C18) bonding and endcapping, bonded to high purity silica substrate.
O Si
O
O
O Si
O
L3
1-5
CH 3
O Si
Polar Group
CH3
HILIC
Low pH = 45 C
High pH = 45 C
3, 5 m
100
330 m2/g
unbonded
Selectivity Features: Excellent for retention of very polar, basic, water soluble analytes. Specifically designed and tested for
HILIC separations using mobile phases containing high concentrations of organic solvent.
Bonding: Unbonded high purity silica substrate.
O
O Si
O
O
OO Si
O OSi
O
Low pH = 45 C
3, 5, 10 m
100
330 m2/g
High pH = 45 C
Selectivity Features: Retention of polar compounds. Designed for compatibility with 100% aqueous mobile phases.
Bonding: Difunctional C18 bonding, fully endcapped, bonded to high purity silica substrate.
L1
C18
OO Si
OSi
O
O
O Si
O
CH 3
O Si
O
3-7
12%
Polar Group
Si 3
O CH
O
CH 3
O Si
O
Polar Group
Si 3
O CH
O
HSS C18
O
OOSi
OO Si
L1
1-8
Low pH = 45 C
High pH = 45 C
3.5, 5 m
100
230 m2/g
15%
Selectivity Features: High performance C18 chemistry, increased retention, superior peak shape, resists acid hydrolysis at low pH.
Designed for UPLC separations where silica-based C18 selectivities are desired.
Bonding: High coverage trifunctional C18, fully endcapped, bonded to High Strength Silica (HSS) HPLC particle substrate.
OO Si
O
CH 3
O Si
CH3
HSS C18 SB
O
OOSi
OO Si
Polar Group
L1
www.waters.com
O
O Si
O
Low pH = 45 C
High pH = 45 C
3.5, 5 m
100
230 m2/g
8%
Selectivity Features: Unique, non-endcapped C18 chemistry designed specifically for method development scientists. Offers unique
Selectivity for Bases (SB) when operating under low pH conditions and transferability between UPLC and HPLC separations.
Bonding: Intermediate coverage tri-functionally bonded C18, no endcapping, bonded to High Strength Silica (HSS) HPLC particle substrate.
L1
HSS T3
2-8
2-8
Low pH = 45 C
High pH = 45 C
3.5, 5 m
100
230 m2/g
11%
Selectivity Features: Aqueous mobile-phase compatible HPLC column designed for extreme retention.
Combines polar compound retention with transferability between UPLC and HPLC separations.
Bonding: T3 (C18) bonding and endcapping, bonded to High Strength Silica (HSS) HPLC particle substrate.
Analytical Chromatography
111
aNaLYTICaL ChROMaTOGRaPhY
Hydrophobicity
(k, Ethylbenzene)
Chelating
(Tailing Factor (TF), Quinizarin)
Silanol Activity
(k, Amitriptyline)
Silanol Activity
(TF, Amitriptyline)
Shape Selectivity
(Bonding Density, mol/m2)
1.63
1.30
2.54
1.09
2.23
1.7
1.87
1.0
2.0
2.54
1.09
2.23
1.5
2.5
2.25
1.65
0.8
2.0
1.63
1.30
1.0
1.5
2.0
1.10
1.36
1.36
1.07
1.19
1.1
0.98
6
2
1.36
1.07
1.19
7.5
1.2
1.7
1.5
no peak
no peak
1.10
1.36
1.2
1.1
no peak
3.79
2.37
5.96
22.2
6.50
5.1
5.32
7.5
6.3
5.96
22.2
6.50
4.6
6.3
5.1
3.1
23
11.5
3.79
2.37
1.7
3.3
35
1.10
1.13
1.41
7.2
3.15
2.4
1.35
4
1.9
1.41
7.2
3.15
3
1.1
1.7
1.2
3
7
1.10
1.13
1.1
1.3
8
3.1
3.3
3.2
1.6
1.6
1.6
1.6
1.1
3.6
3.2
1.6
1.6
2.7
3.5
3.2
3.3
1.7
2.6
3.1
3.3
2.3
2.2
2.8
* For use in C18 column brand name selection when referencing USP monographs which specify the characteristics/performance of the C18 column used in the method.
Choose a brand as close as possible to the referenced column values for the best chance of duplicating the analytical results of the method.
www.waters.com/selectivitychart
112
Analytical Chromatography
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
Brand
Particle Size
AccQTag Ultra
ACQUITY UPLC CSH C18
ACQUITY UPLC BEH C18
ACQUITY UPLC Shield RP18
nanoACQUITY UPLC BEH130
nanoACQUITY UPLC BEH300
ACQUITY UPLC OST C18
ACQUITY UPLC HSS C18
ACQUITY UPLC HSS C18 SB
ACQUITY UPLC HSS T3
Atlantis T3
Atlantis dC18
BioSuite PA-B
BioSuite PA-A
Bondapak C18
Bondapak C18 Radial-Pak
HSS C18
HSS C18 SB
HSS T3
Delta-Pak C18
Nova-Pak C18
Prep Nova-Pak HR C18
Resolve C18
SunFire C18
Symmetry C18
SymmetryPrep C18
Symmetry300 C18
SymmetryShield RP18
Waters Spherisorb ODS1
Waters Spherisorb ODS2
Waters Spherisorb ODSB
XBridge C18
XBridge Shield RP18
XBridge BEH130
XBridge BEH300
XBridge OST C18
XSelect CSH C18
XTerra MS C18
XTerra RP18
L2
1.7
1.7
1.7
1.7
1.7
1.7
1.7
1.8
1.8
1.8
3, 5, 10
3, 5, 10
3
3
10
10
3.5, 5,
3.5, 5,
3.5, 5
5
4, 6
6
5, 10
3.5, 5, 10
3.5, 5, 7
3.5, 5, 7
3.5, 5
3.5, 5
3, 5, 10
3, 5, 10
3, 5, 10
2.5, 3.5, 5, 10
2.5, 3.5, 5, 10
3.5, 5, 10
3.5, 5, 10
2.5
3.5, 5
2.5, 3.5, 5, 10
3.5, 5, 10
Type
Page
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Irregular
Irregular
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
240
91
93
93
262
262
246
95
95
95
133
133
212
212
158
194
138
138
138
159
156
191
161
131
146
190
150
148
153
153
153
127
127
208
208
246
121
142
142
Brand
Bondapak Prep C18
Particle Size
Type
50
Irregular
Page
191
L3
Brand
Particle Size
Type
Page
1.7
3, 5
5, 8
7.5
6, 4
10
5, 10
5, 10
5, 10
2.5, 3.5, 5
Spherical
Spherical
Spherical
Spherical
Spherical
Irregular
Spherical
Spherical
Spherical
Spherical
93
137
233
233
156
160
161
186
153
127
L4
Brand
Porasil Prep Silica
Particle Size
Type
Page
50
Irregular
192
L5
L6
L7
Brand
ACQUITY UPLC BEH C8
Nova-Pak C8
Resolve C8
Waters Spherisorb C8
SunFire C8
Symmetry C8
SymmetryShield RP8
SymmetryPrep C8
XBridge C8
XTerra MS C8
XTerra RP8
L8
Type
Page
1.7
4, 6
5, 10
3, 5, 10
3.5, 5, 10
3.5, 5, 7
3.5, 5
7
2.5, 3.5, 5, 10
2.5, 3.5, 5, 10
3.5, 5, 10
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
93
156
161
153
186
146
148
190
127
142
142
Brand
Bondapak NH2
High Performance Carbohydrate Analysis
Waters Spherisorb NH2
L9
Particle Size
Particle Size
Type
Page
10
3, 5
3, 5, 10
Irregular
158
170
153
Spherical
Brand
Spherisorb SCX
Particle Size
Type
Page
5, 10
Spherical
153
www.waters.com
Analytical Chromatography
113
aNaLYTICaL ChROMaTOGRaPhY
L10
Brand
Bondapak CN
Nova-Pak CN HP
Resolve CN
Waters Spherisorb CN
L11
L19
Type
Page
10
4
5, 10
3, 5, 10
Irregular
Spherical
Spherical
Spherical
158
156
161
153
Brand
Particle Size
Type
Page
1.7
1.7
10
4
2.5, 3.5, 5
3, 5, 10
3.5, 5
3.5, 5
Spherical
Spherical
Irregular
Spherical
Spherical
Spherical
Spherical
Spherical
99
99
158
156
127
153
121
141
L12
Particle Size
AccellPlus QMA
L13
Waters Spherisorb C1
Irregular
230
Particle Size
Type
3, 5, 10
Spherical
Page
154
Particle Size
Type
5, 10
Spherical
Page
154
Brand
Waters Spherisorb C6
Particle Size
Type
3, 5, 10
Spherical
Page
154
L16
L17
Brand
Fast Fruit Juice
IC-Pak Ion Exclusion
IC-Pak Cation
Shodex RSpak DC-613
L18
Particle Size
Type
Page
N/A
7
10
(6)
N/A
Spherical
Spherical
Spherical
173
174
175
162
Particle Size
Type
Page
9
(7)
Spherical
Spherical
170
170
Brand
Particle Size
Type
Page
4, 5, 8, 10,
13, 17
Spherical
233
N/A
Irregular
Irregular
Irregular
Irregular
Irregular
Irregular
213
234
234
234
234
234
234
Particle Size
Type
Page
Spherical
Spherical
Spherical
Spherical
162
272
272
272
Insulin HMW P
Protein-Pak 60
Protein Pak 125
Protein-Pak 300SW
Protein-Pak KW -802.5
Protein-Pak KW -803
Protein-Pak KW -804
L22
Brand
IC-Pak Ion Exclusion
Shodex RSpak DC 613
Shodex SP-0810
L23
10
10
10
7
7
7
Brand
Page
Brand
L15
Type
Brand
L14
50
Sugar-Pak 1
Shodex SC-1011
L21
Brand
Brand
L20
Particle Size
Type
Page
7
6
8
Spherical
Spherical
Spherical
175
162
170
Brand
Particle Size
Type
Page
BioSuite Q AXC
BioSuite DEAE
BioSuite Q-PEEK
IC-Pak Anion
Protein-Pak Q 8HR
10, 13
2.5, 10, 13
10
10
8
Spherical
Spherical
Spherical
Spherical
Spherical
226
226
226
174
227
L24
L25
Brand
Ultrahydrogel DP, + 120
Particle Size
Type
Page
10
Spherical
279
114
Analytical Chromatography
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
L26
Brand
Particle Size
Type
Page
1.7
Spherical
Spherical
[100+300]
Spherical
Spherical
222
Symmetry300 C4
XBridge BEH300 C4
L27
L37
3.5
3.5
160
150
222
Brand
Ultrahydrogel 250
L38
Ultrahydrogel
Particle Size
Type
Page
Porasil
37-55
Irregular
160
L39
Particle Size
Type
Page
N/A
Spherical
170
Particle Size
Type
Page
N/A
Spherical
170
Particle Size
Type
Page
N/A
Spherical
170
Brand
Brand
L28
A hydrophilic-polyhydroxymethacrylate gel
of totally porous spherical resin.
Brand
Ultrahydrogel
L40
L29
L41
L30
L42
L31
L43
L32
L33
Brand
BEH200 SEC
L34
Particle Size
Type
Page
1.7
Spherical
217
Brand
Shodex SP0810
Particle Size
Type
Page
N/A
Spherical
170
L35
L36
Brand
ACQUITY UPLC CSH Fluoro-Phenyl
XSelect CSH Fluoro-Phenyl
Particle Size
Type
Page
1.7
3.5, 5
Spherical
Spherical
99
99
L44
L45
L46
L55
Brand
Particle Size
Type
IC-Pak C M/D
L59
Page
175
Brand
Biosuite 125, 250, 450 Series
Particle Size
Type
Page
4-17
Spherical
233
www.waters.com
Analytical Chromatography
115
aNaLYTICaL ChROMaTOGRaPhY
Physical Characteristics of HPLC and UPLC Packing Materials
Brand
AccQTag Ultra
ACQUITY UPLC CSH
Chemistry
C18
C18
Phenyl-Hexyl
Fluoro-Phenyl
ACQUITY UPLC BEH
C18
C8
Shield RP18
Phenyl
HILIC
Amide
ACQUITY UPLC BEH200
SEC
nanoACQUITY UPLC BEH130
C18
nanoACQUITY UPLC BEH300
C18
C4
ACQUITY UPLC HSS
T3 C18
C18
C18 SB
Atlantis
T3 C18
dC18
HILIC
Bondapak
C18
Phenyl
CN
NH2
Bondapak
C18
C18
Delta-Pak
C4
C18
C4
C18
HSS
T3 C18
C18
C18 SB
Nova-Pak
C18
C8
Phenyl
CN HP
Silica
Porasil
Silica
Porasil
Silica
Resolve
C18
C8
CN
Silica
SunFire
C18
C8
Silica
Symmetry
C18
C8
SymmetryShield
RP8
RP18
SymmetryPrep
C18
C8
Symmetry300
C18
C4
116
Particle Shape
Particle Size(s)
Pore Size*
% Carbon Load
Endcapped
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Irregular
Irregular
Irregular
Irregular
Irregular
Irregular
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Irregular
Irregular
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.8 m
1.8 m
1.8 m
3, 5, 10 m
3, 5, 10 m
3, 5 m
10 m
10 m
10 m
10 m
15-20 m
15-20 m
5, 15 m
5, 15 m
5, 15 m
5, 15 m
3.5, 5 m
3.5, 5 m
3.5, 5 m
4, 6 m
4 m
4 m
4 m
4, 6 m
10 m
15-20 m
5, 10 m
5 m
10 m
5, 10 m
2.5, 3.5, 5, 10 m
2.5, 3.5, 5, 10 m
5, 10 m
3.5, 5 m
3.5, 5 m
3.5, 5 m
5 m
7 m
7 m
3.5, 5 m
3.5, 5 m
135
130
130
130
130
130
130
130
130
130
200
130
300
300
100
100
100
100
100
100
125
125
125
125
125
300
100
100
300
300
100
100
100
60
60
60
60
60
125
125
90
90
90
90
100
100
100
100
100
100
100
100
100
300
300
185
185
185
185
185
185
185
185
185
185
216
185
90
90
230
230
230
330
330
330
330
330
330
330
330
100
300
300
125
125
230
230
230
120
120
120
120
120
330
330
200
200
200
200
340
340
340
335
335
335
335
335
335
110
110
0.70
0.70
0.70
0.70
0.70
0.70
0.70
0.70
0.70
0.70
high
0.70
0.66
0.66
0.70
0.70
0.70
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
0.70
0.70
0.70
0.30
0.30
0.30
0.30
0.30
1.00
1.00
0.50
0.50
0.50
0.50
0.90
0.90
0.90
0.90
0.90
0.90
0.90
0.90
0.90
0.80
0.80
17
15
14
10
17
13
17
15
unbonded
12
12
17
12
8
11
15
8
14
12
unbonded
9.8
9.3
6
4
10
3.5
7.3
17
2.6
6.8
11
15
8
7.3
4
4.6
3
n/a
n/a
n/a
10.2
5.1
3
n/a
16
12
unbonded
19.1
11.7
15
17
19.1
11.7
8.5
2.8
yes
yes
yes
no
yes
yes
yes
yes
n/a
n/a
n/a
yes
yes
no
yes
yes
no
yes
yes
n/a
yes
yes
yes
no
yes
yes
yes
yes
yes
yes
yes
yes
no
yes
yes
yes
yes
n/a
n/a
n/a
no
no
no
n/a
yes
yes
n/a
yes
yes
yes
yes
yes
yes
yes
yes
Analytical Chromatography
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aNaLYTICaL ChROMaTOGRaPhY
Brand
Waters Spherisorb
XBridge
XBridge BEH130
XBridge BEH300
XTerra
XSelect CSH
Chemistry
Particle Shape
Particle Size(s)
Pore Size*
% Carbon Load
Endcapped
Silica
ODS2
ODS
ODSB
C8
C6
C1
Nitrile
Amino
Phenyl
OD/CN
SAX, SCX
C18
C8
Shield RP18
Phenyl
HILIC
Amide
C18
C18
C4
RP18
RP8
MS C18
MS C8
Phenyl
C18
Phenyl-Hexyl
Fluoro-Phenyl
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
3, 5, 10 m
3, 5, 10 m
3, 5, 10 m
5 m
3, 5, 10 m
3, 5, 10 m
3, 5, 10 m
3, 5, 10 m
3, 5, 10 m
3, 5, 10 m
5 m
5, 10 m
2.5, 3.5, 5, 10 m
2.5, 3.5, 5, 10 m
2.5, 3.5, 5, 10 m
2.5, 3.5, 5 m
2.5, 3.5, 5 m
3.5 m
3.5, 5, 10 m
3.5, 5, 10 m
3.5 m
3.5, 5, 10 m
3.5, 5, 10 m
2.5, 3.5, 5, 10 m
2.5, 3.5, 5, 10 m
3.5, 5 m
3.5, 5 m
3.5, 5 m
3.5, 5 m
80
80
80
80
80
80
80
80
80
80
80
80
135
135
135
135
130
130
135
300
300
125
125
125
125
125
135
135
135
220
220
220
220
220
220
220
220
220
220
220
220
185
185
185
185
185
185
185
86
90
175
175
175
175
175
185
185
185
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.70
0.70
0.70
0.70
0.70
0.70
0.70
0.66
0.66
0.70
0.70
0.70
0.70
0.70
0.70
0.70
0.70
n/a
11.5
6.2
11.5
5.8
4.7
2.2
3.1
1.9
2.5
5
4
17.5
17.5
17.5
17.5
unbonded
12
17.5
12
8
15
13.5
15.5
12
12
15
14
10
n/a
yes
no
yes
yes
yes
no
no
no
no
yes
no
yes
yes
yes
yes
n/a
n/a
yes
yes
no
yes
yes
yes
yes
yes
yes
yes
no
* Nominal value
Troubleshooting
Description
Part No.
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Analytical Chromatography
WAT038216
117
aNaLYTICaL ChROMaTOGRaPhY
Ligand Type
Fluoro-Phenyl
F
O Si
O
O Si
O
Phenyl-Hexyl
O
F
O Si
O
Trifunctional C18
Trifunctional Propylfluorophenyl
Trifunctional C6 Phenyl
3.5, 5 m
3.5, 5 m
3.5, 5 m
2.3 mol/m2
2.3 mol/m2
2.3 mol/m2
Carbon Load*
15
10
14
End-cap Style
Proprietary
None
Proprietary
1-11
1- 8
1-11
Low pH=80 C
Low pH=60 C
Low pH=80 C
High pH=45 C
High pH=45 C
High pH=45 C
pH Range
Temperature Limits
* Expected or Approximate Values
118
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aNaLYTICaL ChROMaTOGRaPhY
Engineered to Provide a Wider
Range of Selectivity
Reproducibility
10
6
7
1-R2
15
CSH C18
XSelect CSH Fluoro-Phenyl 3.5 m
CSH Fluoro-Phenyl
CSH Phenyl-Hexyl
10
R2 = 0.76
S = 49
0
0
10
15
kg on XBridge C18
15
BEH RP18
kg on XBridge Phases
BEH Phenyl
BEH C8
10
0.0
R2 = 0.97
S = 16
0
10
15
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0 min
ACQUITY UPLC CSH Fluoro-Phenyl (1.7 m) and XSelect CSH Fluoro-Phenyl (3.5 and 5 m)
reproducibility and scalability. Gradient separations on 2.1 x 50 mm columns containing nine different
batches of CSH Technology Fluoro-Phenyl representing three (1.7, 3.5, and 5 m) paricle sizes. Gradient:
A:15.4mMammonium formate, pH 3; B: acetonitrile; 5 to 90% B linear in 5 minutes. Temperature:
30 C. Injection volume: 5 L. Detection: 254 nm. Flow rate: 0.5 mL/min. Analytes: (1) thiourea;
(2) resorcinol; (3)metoprolol; (4) 3-nitrophenol; (5) 2-chlorobenzoic acid; (6) amitriptyline;
(7) diethylphthalate; (8)fenoprofen; (9) dipropylphthalate; (10) pyrenesulfonic acid. System: ACQUITY UPLC.
kg on XBridge C18
Plots of gradient retention factors (kg) on XSelect (top) or XBridge (bottom) columns versus
gradient retention factors on an XBridge C18 column. T he more scatter around the regression line,
the larger the selectivity differences.
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119
aNaLYTICaL ChROMaTOGRaPhY
TFA Performance without TFA Suppression
CSH Technology provides TFA peak shape performance without the use of
TFA (and its associated disadvantages). Improved peak shape performance is
realized with a more MS-friendly mobile-phase additive such as formic acid
(e.g., 0.1% HCOOH). Further, because formic acid does not ex hibit
the lingering c hromatographic effects of an ion-pairing agent, method
development is faster and more straightforward.
120
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ANALYTICAL CHROMATOGRAPHY
XSelect Analytical Columns
Dimension
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
2.1 x 10 mm
2.1 x 20 mm IS
2.1 x 30 mm
2.1 x 50 mm
2.1 x 100 mm
2.1 x 75 mm
2.1 x 150 mm
3.0 x 20 mm
3.0 x 20 mm IS
3.0 x 30 mm
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
4.6 x 20 mm
4.6 x 20 mm IS
4.6 x 30 mm
4.6 x 50 mm
4.6 x 75 mm
4.6 x 100 mm
4.6 x 150 mm
2.1 x 10 mm
2.1 x 20 mm IS
2.1 x 30 mm
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 20 mm
3.0 x 20 mm IS
3.0 x 30 mm
3.0 x 50 mm
3.0 x 100 mm
3.0 x 150 mm
3.0 x 250 mm
4.6 x 20 mm IS
4.6 x 20 mm
4.6 x 30 mm
4.6 x 50 mm
4.6 x 75 mm
4.6 x 100 mm
4.6 x 150 mm
4.6 x 250 mm
Type
Particle Size
C18
Fluoro-Phenyl
Phenyl-Hexyl
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
186005249
186005250
186005251
1860052521
186005253
186005254
186005255
186005256
186005644
186005257
1860052582
186005259
186005260
186005261
186005647
186005262
186005263
1860052642
186005265
186005266
186005267
186005268
186005269
186005270
1860052711
186005272
186005273
186005274
186005275
186005276
1860052772
186005278
186005279
186005280
186005281
186005282
186005283
186005284
1860052852
186005286
186005287
186005288
186005289
186005290
186005291
186005304
186005305
186005306
1860053071
186005308
186005309
186005310
186005311
186005646
186005312
1860053132
186005314
186005315
186005316
186005649
186005317
186005318
1860053192
186005320
186005321
186005322
186005323
186005324
186005325
1860053261
186005327
186005328
186005329
186005330
186005331
1860053322
186005333
186005334
186005335
186005336
186005337
186005338
186005339
1860053402
186005341
186005342
186005343
186005344
186005345
186005346
186005359
186005360
186005361
1860053621
186005363
186005364
186005365
186005366
186005645
186005367
1860053682
186005369
186005370
186005371
186005648
186005372
186005373
1860053742
186005375
186005376
186005377
186005378
186005379
186005380
1860053811
186005382
186005383
186005384
186005385
186005386
1860053872
186005388
186005389
186005390
186005391
186005392
186005393
186005394
1860053952
186005396
186005397
186005398
186005399
186005400
186005401
Particle Size
C18
Fluoro-Phenyl
Phenyl-Hexyl
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
5 m
5 m
5 m
186005538
186005539
186005540
186005541
186005542
186005543
186005544
186005545
186005546
186005547
186005548
186005549
186005550
186005551
186005552
186005553
186005554
186005555
186005556
186005557
186005558
186005559
186005560
186005561
186005562
186005563
186005564
186005565
186005566
186005567
186005568
186005569
186005570
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121
aNaLYTICaL ChROMaTOGRaPhY
XBridge Columns
A New Milestone in Chromatography
One of the most important parameters in the design of the new hybrid
particle was to significantly improve the chromatographic equivalence to
state-of-the-art silica based materials. T he origins of band spreading, which
decreases separation efficiency, are described by the van Deemter equation:
Data for both silica and BEH Technology packings, fitted to the van Deemter
equation demonstrate effective equivalence in efficiency. The c term for
XBridge C18 is virtually identical to that for the two state-of-the-art C18
silica columns, indicating that all these columns are comparable in mass
transfer characteristics.
h = a + b/u + cu
BEH Technology
Analyte:
Mobile Phase:
Temperature:
12
10
Maximize efficiency
Decanophenone
70/30 ACN/water
30 C
XBridge C18 5m
Symmetry C18 5m
SunFire C18 5m
1.04
0.76
0.95
8.7
10.6
11.6
0.044
0.045
0.040
r2
0.998
0.999
0.997
20
40
60
80
u [uidp/Dm]
The reduced plate height, h, is a function of the reduced linear velocity, u, [both normalized for particle size]
and a, b, and c summarize the contributions of eddy diffusion, longitudinal diffusion, and the sum of
stationary- and mobile-phase mass transfer terms, respectively.
* Patent # 6,686,035 B2
122
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aNaLYTICaL ChROMaTOGRaPhY
pH Stability
Improved Low pH Performance
XBridge packings incorporate the use of well-characterized, state-of-theart, proprietary procedures for bonding and endcapping. Bonded phases
prepared using these procedures are much more stable and reproducible
at low pH than conventionally prepared materials and resist the typical
ligand hydrolysis failure. In an accelerated low pH stability test, XBridge
C18 columns show very little retention loss and exhibit a column lifetime
equivalent to that of a sterically hindered C18 silica-bonded phase.
Probe: Methylparaben
XBridge C18
Zorbax SB-C18
XBridge Phenyl
80
Zorbax SB Phenyl
90
% Initial N5s
90
% Initial Retention
Analyte: Acenaphthene
110
100
XBridge C18
XTerra MS C18
Gemini C18
70
Luna C18(2)
Luna C18(2)
70
50
30
0
50
50
50
100
100
200 hours
Compounds
1. Uracil
2. Propranolol
3. Naphthalene
4. Acenaphthene
5. Amitriptyline
5
N=13600
Initial
N=13500
100 h
N=14100
200 h
N=11700
300 h
pH 3
Desipramine
XBridge Family
Amitriptyline
pH 7
Nortriptyline
Amitriptyline
150
Capcell PAK MG II
C8
Phenyl
RP18
C18
Zorbax
SB-C18
12
16
20
24 Min.
Chromatograms, run at regular intervals during the high-pH lifetime study, verify
that 86% of the original XBridge column efficiency remains after 300 hours at pH 10
and elevated temperature, with little change in peak shape or retention time.
The combination of excellent particle and ligand stability as well as high chromatographic
efficiencies makes XBridge columns an ideal choice for low and intermediate pH methods.
www.waters.com
123
aNaLYTICaL ChROMaTOGRaPhY
Column Lifetime and Buffer Choice
30 Hours
300 Hours
N = 2218
N = 11700
2
5
2
3
3
12
16
20
24 Min.
10
20
30 Hours
13 Hours
300 Hours
27050
Hours
mM TEA, 50 C, pH 10
12
10
16
15
20
24 Min.
20 Min.
0
0
10
10
15
20 Min.
5
10
15
Phenomenex
Gemini
C18
10
15
30 mM KH2PO4, 30 C, pH 12.3
5
6
10
11
11
7,8
9
13
9,10
12
12
9,10
78
5
11
13
12
13
11
2
2
12
12
7
7
10
5
10
5
13
78
12
12
16
20 Min.
12
16
20 Min.
12
16
20 Min.
12
16
20 Min.
2
2
6
6
4
4
4,5
4,5
10
11
10
11
67
12
12
13
10
5
10
11,12
6
2
2
13
3
5
10
13
9
9
12
11,12
10
10
11
Phenyl
Phenyl
11
12
86 7
11
13
4
1
7
10
12
13
9
9
11
Shield RP18
2
Shield RP18
12
12
12
3
3
6
7
2
1
11
5 6
Phenyl
Phenyl
12
23
11
6
12
Shield RP18
Shield RP18
2
11
11
9,10
9,10
7
10
10
13
8
8
12
16
20 Min.
12
16
20 Min.
12
16
20 Min.
12
16
20 Min.
Literature
References
Waters XBridge HPLC
Columns Brochure,
Literature Reference 720001255EN
A Review of Waters Hybrid Particle Technology
Part 2. Ethylene-Bridged (BEH TEchnology)
Hybrids and T heir Use in Liquid
Chromatography W hite Paper,
Literature Reference 720001159EN
124
11
Phenyl
Phenyl
9
5
11
2
2
10
7,8
12
8,9
8,13
7
Shield RP18
Shield RP18
12
12
11
11
8,13
10
10
12
1
10
10
8,9
20 Min.
3
3
Phenyl
Phenyl
13 Hours
5
6
11
11
16.68
1,2
1,2
30 Min.
20 Min.
11 16.68
20
270 Hours
Shield RP18
Shield RP18
N = 2218
N = 11700
0
0
30 Min.
10
C18
C18
11
5 6
10
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aNaLYTICaL ChROMaTOGRaPhY
XBridge and Phosphate Buffer
With the introduction, and subsequent widespread adoption, of high purity
reversed-phase silica columns in the early 1990s, the method development chemist has been unable to freely utilize phosphate buffers (pH 7
and above) due to their extremely aggressive nature. The combination of
phosphate, intermediate pH and relatively low temperatures (40 C) is well
recognized to lead to short column lifetimes for most modern phases.
Phosphate buffers however do have quite desirable qualities for a non-MS
based assay:
Excellent UV transparency
Mobile Phase B:
Flow Rate:
Gradient:
Injection Volume:
Sample Concentration:
Temperature:
Detection:
Instrument:
Mobile Phase A:
Mobile Phase B:
Isocratic:
Compounds
1. 1-methylxanthene
2. 1,3-dimethyluric acid
3. theobromine
4. 1,7-dimethylxanthene
0.1% FA in H2O
0.1% FA in ACN
95% A: 5% B
5 m-150 mm
Injection = 5.0 L
Rs(2,3) = 2.28
Flow Rate: 0.2 mL/min
4
Compounds
1. Doxylamine
2. Benzamide
3. Hydroxyisophthalic Acid
4. Doxepine
5. Flavone
6. Fenoprofen
10 Min.
3.5 m-100 mm
Injection = 3.3 L
Rs(2,3) = 2.32
Flow Rate: 0.3 mL/min
HPLC
4 Min.
2.5 m-75 mm
Injection = 2.5 L
Rs(2,3) = 2.34
Flow Rate: 0.5 mL/min
pH 2
0.4
0.8
1.2
1.6
2 Min.
1.7 m-50 mm
Injection = 1.7 L
Rs(2,3) = 2.29
Flow Rate: 0.6 mL/min
pH 7
UPLC
0
2
6
pH 12
0.2
0.4
0.6
0.8
1.10 Min.
8 Min.
XBridge columns are unique in their ability to withstand these aggressive conditions
(even at pH 12) allowing complete flexibility for the method development chemist,
while maintaining the highest efficiency values associated with silica columns.
www.waters.com
125
ANALYTICAL CHROMATOGRAPHY
XBridge Amide Columns
Since 2003, Waters has developed innovative stationary phases for
Hydrophilic Interaction Chromatography [HILIC] to overcome the challenge
of retaining and separating extremely polar compounds. Based on Waters
novel ethylene bridged hybrid [BEH] particle tec hnology, XBridge
Amide columns utilize a chemically stable, trifunctionally-bonded amide
phase, enabling a new dimension in stability and versatility for HILIC
separations. XBridge Amide columns offer the same selectivity as ACQUITY
UPLC BEH Amide columns, allowing scientists to transfer their separations
between HPLC and UPLC technology platforms.
In addition to enhanced retention of polar compounds, XBridge Amide
columns provide increased mass spectrometry response, direct compatibility
with sample preparation eluates [ PPT, LLE and SPE] as well as orthogonal
selectivity compared to reversed-phase materials, making HILIC an attractive alternative separation technique.
Carbohydrate Analysis
One of the most abundant and diverse classes of compounds analyzed by
HILIC are carbohydrates [monosaccharides, disaccharides, oligosaccharides
and polysaccharides].
T he XBridge Amide column is an exceptional tool for the analysis of
carbohydrates, providing the following benefits:
Inj. Vol.:
Sample Diluent:
Column Temp.:
Needle Wash:
Instrument:
Ionization Mode:
Capillary:
Cone Voltage:
Source Temp.:
Desolv. Temp.:
Desolv. Gas Flow:
SIR m/z:
Ginsenoside Rb1
Orento extract
Dwell Time:
Compounds:
1. Fructose
2. Glucose
3. Sucrose
4. Maltose
5. Maltotriose
6. Maltotetraose
7. Maltopentaose
8. Maltohexa hose
9. Maltoheptaose
Ginsenoside Rb1
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5 min
10 g/mL
10 g/mL
10 g/mL
10 g/mL
10 g/mL
10 g/mL
10 g/mL
10 g/mL
10 g/mL
4
6
2
7
126
10
12
14
16
18
20
22
24
26
28
30
32 min
www.waters.com
Si
Si
3
N
Lin
ke
r
Si
O
O
2.5, 3.5, 5 m
2.5, 3.5, 5 m
3.5 m
3.1 mol/m2
3.2 mol/m2
3.3 mol/m2
3.0 mol/m2
N/A
7.5 mol/m2
Carbon Load*
18%
13%
17%
15%
unbonded
12%
Endcap Style
Si
2.5, 3.5, 5, 10 m
O
Bonded Phase
Si
2.5, 3.5, 5, 10 m
2.5, 3.5, 5, 10 m
Ligand Density*
BEH Particle
Amide
Unbonded
BEH
Trifunctional
C6 Phenyl
Si
Monofunctional
Embedded Polar Group
H
C
Si
Si
3
Si
H
C
O
Po
la
rG
ro
up
Trifunctional C8
Si
Trifunctional C18
Po
la
rG
ro
up
O
O
Si
O
O
Ligand Type
Si
Si
Po
la
rG
ro
up
Si
Si
O
Po
la
rG
ro
up
Si
Si
ANALYTICAL CHROMATOGRAPHY
Proprietary
Proprietary
TMS
Proprietary
N/A
None
pH Range
1-12
1-12
2-11
1-12
1-9
2-11
80 C
60 C
50 C
80 C
45 C
90 C
60 C
60 C
45 C
60 C
45 C
90 C
Pore Diameter*
130
130
130
130
130
130
Pore Volume*
0.7 mL/g
0.7 mL/g
0.7 mL/g
0.7 mL/g
0.7 mL/g
0.7 mL/g
Surface Area*
185 m2/g
185 m2/g
185 m2/g
185 m2/g
185 m2/g
185 m2/g
* Nominal Values
Type
Particle Size
C18
C8
Shield RP18
Phenyl
HILIC
Amide
Column
Guard
Column
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Guard
Column
Column
Column
Column
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
186003118
1860030561
186003201
186003084
186003085
186005626
186003087
1860030572
186003121
186003122
186005630
186003088
1860030582
186003089
186003090
186003124
186003126
186003127
186003128
1860030591
186003019
186003020
186003021
186003022
186003023
186003024
1860030602
186003025
186003026
186003027
186003028
186003164
1860030741
186003167
186003099
186003101
186005627
186003168
1860030752
186003169
186003170
186005631
186003172
1860030762
186003173
186003174
186003175
186003177
186003178
186003179
1860030771
186003180
186003046
186003047
186003048
186003049
186003181
1860030782
186003182
186003050
186003051
186003052
186003136
1860030651
186003139
186003091
186003092
186005628
186003140
1860030662
186003141
186003142
186005632
186003144
1860030672
186003145
186003096
186003146
186003148
186003149
186003150
1860030681
186003151
186003035
186003036
186003037
186003038
186003152
1860030692
186003153
186003039
186003040
186003041
186003306
1860033591
186003307
186003308
186003309
186005629
186003310
1860033602
186003311
186003312
186005633
186003313
1860033612
186003314
186003315
186003316
186003317
186003318
186003319
1860033621
186003320
186003321
186003322
186003323
186003324
186003325
1860033632
186003326
186003327
186003328
186003329
186004455
186004456
186004457
186004458
1860044592
186004460
186004461
186004429
1860044301
186004431
186004432
186004433
186004434
186004435
186004436
186004871
1860048571
186004858
186004859
186004860
186004861
186004862
186004863
186004864
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127
ANALYTICAL CHROMATOGRAPHY
XBridge Analytical Columns
Dimension
4.6 x 20 mm IS
4.6 x 20 mm
4.6 x 30 mm
4.6 x 50 mm
4.6 x 75 mm
4.6 x 100 mm
4.6 x 150 mm
4.6 x 250 mm
2.1 x 10 mm
2.1 x 20 mm IS
2.1 x 30 mm
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 20 mm IS
3.0 x 20 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 100 mm
3.0 x 150 mm
3.0 x 250 mm
4.6 x 20 mm IS
4.6 x 20 mm
4.6 x 30 mm
4.6 x 50 mm
4.6 x 75 mm
4.6 x 100 mm
4.6 x 150 mm
4.6 x 250 mm
1
2
Type
Particle Size
C18
C8
Shield RP18
Phenyl
HILIC
Amide
Column
Guard
Column
Column
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
186003029
1860030612
186003030
186003031
186003032
186003033
186003034
186003943
1860030621
186003107
186003129
186003108
186003109
186003110
186003130
1860030632
186003111
186003131
186003132
186003112
186003133
186003134
1860030642
186003135
186003113
186003114
186003115
186003116
186003117
186003183
1860030792
186003184
186003053
186003185
186003054
186003055
186003963
1860030801
186003186
186003187
186003011
186003012
186003013
186003188
1860030812
186003189
186003190
186003191
186003014
186003192
186003193
1860030822
186003194
186003015
186003195
186003016
186003017
186003018
186003154
1860030702
186003155
186003042
186003043
186003044
186003045
186003964
1860030711
186003156
186003157
186002999
186003002
186003003
186003158
1860030722
186003159
186003160
186003004
186003005
186003161
186003162
1860030732
186003163
186003006
186003007
186003008
186003009
186003010
186003330
1860033642
186003331
186003332
186003333
186003334
186003335
186003965
1860033661
186003336
186003337
186003338
186003339
186003340
186003341
1860033672
186003342
186003343
186003344
186003345
186003346
186003347
1860033682
186003348
186003349
186003350
186003351
186003352
186003353
1860044372
186004438
186004439
186004440
186004441
1860044421
186004443
186004444
186004445
186004446
186004447
186004448
1860044492
186004450
186004451
186004452
186004453
186004454
1860048652
186004866
186004867
186004868
186004869
186004870
128
Type
Particle Size
C18
C8
Shield RP18
Phenyl
MV Kit
MV Kit
MV Kit
MV Kit
MV Kit
MV Kit
MV Kit
MV Kit
MV Kit
MV Kit
MV Kit
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
5 m
5 m
5 m
186003766
186003767
186003768
186003769
186003770
186003771
186003772
186003773
186003774
186003775
186003776
186003777
186003778
186003779
186003780
186003781
186003782
186003783
186003784
186003785
186003786
186003787
186003788
186003789
186003790
186003791
186003792
186003793
186003794
186003795
186003796
186003797
186003798
186003799
186003800
186003801
186003802
186003803
186003804
186003805
186003806
186003807
186003808
186003809
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
90
SunFire C18
2
3
80
SunFire C18
TUSP 1.26
70
Compounds:
1. Uracil
2. Propranolol
3. Butylparaben
4. Naphthalene
5. Acenaphthene
6. Amitriptyline
TUSP 1.35
60
50
0
20
40
60
80
TUSP 1.79
10
15
20
25
30
35
40 min
High Efficiency
A combination of proprietary, state-of-the-art silica synthesis, bonding,
end-capping, and packing technologies produce SunFire columns with high
efficiency. One important benefit of these efforts is greater sensitivity.
MS-Compatibility
SunFire columns are compatible with mass spectrometry applications,
providing sharp peaks, excellent sensitivity, high peak capacity, and
ultra-low bleed. In addition, the speed, excellent resolution, and low
backpressure offered by SunFire Intelligent Speed (IS) columns reduce
costs and analysis times.
1.5
2.0
2.5
3.0
USP Tailing Factor (TUSP)
3.5
4.0
In this comparison test of columns packed with 5 m particles, SunFire C18 column has the lowest USP tailing
factor for the basic probe amitriptyline under the most demanding pH 7 mobile-phase conditions.
www.waters.com
129
aNaLYTICaL ChROMaTOGRaPhY
Batch-to-Batch Reproducibility
product. SunFire columns have demonstrated superior reproducibility
over many years. Batc hes randomly selected over the past 4 years show
excellent reproducibility in the example below.
10
15
20
25
30
35
40 Min.
T his excellent reproducibility is a result of our commitment to maintaining the tightest specifications in the HPLC column industry. SunFire columns start with high purity raw materials,
and are produced using tightly controlled manufacturing processes and column packing procedures that provide todays scientists with the best, most reproducible HPLC columns available.
Min
0.0
4.0
5.0
8.0
3
1
3
1
P = 880 psi
Pc = 49
Rs2/3 = 3.6
130
%B
15
60
15
15
Columns:
%C
10
10
10
10
3
4
%A
75
30
75
75
0.5
1.0
1.5
2.0
2.5
3.0
3.5
0.5
1.0
1.5
2.0
2.5
3.0
3.5
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
Min 4.0%A
0.0
75
6.0 4.030
7.5
75
12.0 75
5.0 Min.
4.5
%B
4.5
15
4.5
60
15
15
%C
5.0
10
10
5.0
10
10
Min.
Min.
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
7.0
7.5 Min.
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
7.0
7.5 Min.
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
7.0
Min %A
0.0
75
8.0
30
10.0 8.075
16.0 75
%B
15
60
15
9.0
15
Injection Volumes:
Column Temperature:
Detection:
Sampling Rate:
Instrument:
modifications)
%C 7.5 Min.
10
10
10
10.0
10
Min.
1.0
2.0
3.0
4.0
5.0
6.0
7.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
9.0
10.0 Min.
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
9.0
10.0 Min.
Mobile Phase A:
Mobile Phase B:
Mobile Phase C:
Flow Rate:
Sample:
www.waters.com
ANALYTICAL CHROMATOGRAPHY
Physical Characteristics
Chemistry
Particle Size
Particle Shape
Pore Size
Carbon Load
End-capped
C18
2.5, 3.5, 5, 10 m
Spherical
100
16%
Yes
C8
2.5, 3.5, 5, 10 m
Spherical
100
12%
Yes
Silica
5, 10 m
Spherical
100
unbonded
N/A
Particle Size
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
2.5 m
C8
186003392
186003397
186003399
186003401
186005634
186003403
186003407
186003409
186005636
186003411
186003415
186003417
186003419
186003394
186003398
186003400
186003402
186005635
186003404
186003408
186003410
186005637
186003412
186003416
186003418
186003420
Particle Size
C18
C8
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
186002526
186002528
186002531
186002532
186002533
186002534
186002535
186002686
186003254
186002542
186002543
186002544
186002549
186002550
186002551
186002552
186002553
186002554
186002705
186002706
186002697
186002709
186002710
186002711
186002712
186002701
Custom
186002719
186002720
186002721
186002699
186002728
186002729
186002730
186002731
186002732
Dimension
1.0 x 150 mm
2.1 x 20 mm IS
2.1 x 30 mm
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 20 mm IS
3.0 x 50 mm
3.0 x 100 mm
3.0 x 150 mm
3.0 x 250 mm
4.6 x 20 mm IS
4.6 x 30 mm
4.6 x 50 mm
4.6 x 100 mm
4.6 x 150 mm
4.6 x 250 mm
Dimension
www.waters.com
Particle Size
186002529
186002537
186002538
186002539
186002540
186002541
186002685
186002545
186002546
186002547
186002548
186002555
186002556
186002557
186002558
186002559
186002560
C8
186002707
186002698
186002714
186002715
186002716
186002717
186002702
186002723
186002724
186002725
186002726
186002700
186002734
186002735
186002736
186002737
186002738
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
5 m
C18
186002674
186002675
186002676
186002677
186002678
186002679
186002680
C8
186002739
186002740
186002741
186002742
186002743
186002744
186002745
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
C18
Literature
References
SunFire Columns-T he Standard in Silica-Based
HPLC Column Performance Brochure,
Literature Reference 720000875EN
Particle Size
3
4
Particle Size
C18
C8
2.5 m
2.5 m
2.5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
1860033953
1860033963
1860034064
1860034144
1860027083
1860027184
1860027274
1860027133
1860027334
1860027224
1860034054
1860034134
1860025303
1860026814
1860026824
1860025363
1860026844
1860026834
Requires 2.1 x 10 mm Universal Sentry Guard Column Holder, Part No. WAT097958
Requires 3.0 x 20 mm Universal Sentry Guard Column Holder, Part No. WAT046910
131
aNaLYTICaL ChROMaTOGRaPhY
Atlantis Columns
T he Atlantis family of HPLC columns was created to solve one of the
most challenging chromatographic problems, retaining polar compounds.
T hrough historical experience and synthetic innovation, Atlantis columns
lead t he industry in providing exceptional performance, versatility,
retention and chromatographic stability for polar compounds, while also
affording balanced retention for broad analyte mixtures.
For retention and separation of polar compounds via reversed-phase HPLC,
Atlantis T3 columns set the standard for polar compound retention. Atlantis
T3 columns are a universal, silica-based, reversed-phase C18 line of HPLC
columns that not only retain and separate small, water-soluble polar
organic compounds, but also provide superior performance across a wider
pH range. Atlantis columns are the culmination of over 30 years of bonded
phase researc h and are a vast improvement upon the highly successful
Base
Atlantis T3
or
XBridge C18
Yes
Is the polar
compound an
acid, neutral
or base?
Can it be
retained by
reversed-phase?
Can it be
retained by
reversed-phase?
No
Yes
Sufficient mass
spectrometry
sensitivity?
Acid or Neutral
Yes
Atlantis T3
No
Insufficient
retention?
Yes
No
Key:
Atlantis T3 = low pH (pH 3), high aqueous (>95%)
XBridge C18 = high pH (pH10) high aqueous (>95%)
Alternate separation
technique, such as
ion-pair or
ion-exchange
Atlantis HILIC Silica = low pH (pH 3), 10 mM ammonium formate with 0.2% formic acid in 95% ACN:5% H2O
XBridge HILIC = mid pH (pH 5), 10 mM ammonium acetate with 0.02% acetic acid in 95% ACN:5% H2O
132
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
T3 Bonding
T he superior performance of Atlantis T3 HPLC columns and the ACQUITY
UPLC HSS T3 columns is a result of Waters advanced T3 bonding process.
T3 bonding utilizes a trifunctional C18 alkyl phase bonded at a ligand
density that promotes polar compound retention and aqueous mobile-phase
compatibility. T he proprietary T3 endcapping process is muc h more
effective than traditional trimethyl-silane (TMS) endcapping.
4. Guanosine - 5- monophosphate
5. T hymine
5
Atlantis T3
3,5
Agilent Zorbax SB-Aq
5
Thermo Aquasil C18
3
1
2,4
3,4
5
Phenomenex Synergi Hydro-RP
9 Min.
Atlantis T3
SunFire C18
Phenomenex Synergi Hydro-RP
Shiseido Capcell Pak C18 AQ
Phenomenex Synergi Polar-RP
Agilent Zorbax Eclipse Plus
Atlantis dC18
Phenomenex Aqua
GL Sciences Inertsil ODS-SP
10
20
30
40
50
60
70%
www.waters.com
133
aNaLYTICaL ChROMaTOGRaPhY
Improved pH 7 Performance
At pH 7, poor peak shape for amine-containing bases and shortened
column lifetimes are encountered when using intermediate ligand density
C18 columns designed for polar compound retention. Poor peak shape is due
to secondary interactions with unreacted silanols that remain present after
bonding and endcapping. T he proprietary T3 endcapping procedure reacts
with more of these active silanols thereby dramatically improving peak
shape for bases. Shortened column lifetimes are due to the dissolution of
the silica particle substrate by the high pH mobile phase. T he more effective
and efficient T3 bonding helps protect the silica substrate from dissolution,
thus providing longer column lifetimes.
Atlantis T3
GL Sciences Inertsil ODS-SP
Shiseido Capcell PAK C18 AQ
Phenomenex Synergi Polar-RP
Phenomenex Synergi Hydro-RP
Phenomenex Aqua
Atlantis dC18
Agilent Zorbax SB-Aq
T hermo Aquasil C18
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
Injection Volume:
Column Temp.:
Detection:
%D
17.5
27.5
41
41
17.5
17.5
Compounds:
1. Propyl gallate
2. 2,4,5-Trihydroxybutyrophenone
3. Tert-butylhydroquinone
4. Butylated hydroxyanisole
5. 2,6-Di-tert-butyl-4-hydroxymethylphenol
6. Octyl gallate
7. Butylated hydroxytoluene
8. Lauryl gallate
4
1
2
5
134
10
12
14 Min.
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
ACQUITY UPLC HSS T3 Columns for Polar Compound Retention
ACQUIT Y UPLC HSS (High Strength Silica) columns contain the first
and only 100% silica particle designed, tested and intended for use
in applications up to 15000 psi/1000 bar. T he ACQUIT Y UPLC HSS
particle is not an HPLC particle. High pore volume HPLC particles do not
posses the mechanical stability necessary to withstand the high column
packing and operating pressures of UPLC separations.
Compounds
1. Norepinephrine
2. Epinephrine
3. Dopamine
4. 3,4- Dihydroxyphenylacetic acid (DOPAC)
5. Serotonin (5-HT)
6. 5-Hydroxy-3-indoleacetic acid (5-HIAA)
7
7. 4-Hydroxy-3-methoxyphenylacetic acid (HVA)
Mobile Phase A:
Mobile Phase B:
Flow Rate:
Isocratic1Mobile
3
Phase Composition:
2
Injection Volume:
Temperature:
Detection:
0
Instrument:
2% B
0.7 L
30 C
UV @ 280 nm
1
ACQUITY UPLC System with
ACQUITY UPLC 2996 PDA
4 Min.
5
1
4 Min.
1
3
6
4
2
3
1
UPLC Separation on
1 UPLC
ACQUITY
1
2
3
4HSS5 T3 6
0.2
4
5 n Tim e
80% Ru
ction!!
du
Re
0.6
0.8
1.0
0.4
9 Min.
1.4 Min
1.2
1
6
2
3
0
0.2
0.4
1. Cytosine
2.77
2. 5-Fluorocytosine
3.58
4.56
3. Uracil
3.93
1.71
4. 5-Fluorouracil
4.59
3.41
5. Guanine
5.92
7.76
6. T hymine
7.63
11.01
7. Adenine
8.39
4.70
Similar Selectivities
USP
Resolution
Retention
Time (min)
9 Min.
Compound
Compound
Retention
Time (min)
USP
Resolution
1. Cytosine
0.34
2. 5-Fluorocytosine
0.44
5.02
3. Uracil
0.48
1.50
4. 5-Fluorouracil
0.56
3.17
5. Guanine
0.80
8.16
6. T hymine
1.25
14.65
7. Adenine
1.33
3.50
5
0.6
0.8
1.0
1.2
1.4 Min
www.waters.com
135
aNaLYTICaL ChROMaTOGRaPhY
Atlantis HILIC Silica
Hydrophilic-Interaction
Chromatography (HILIC)
HILIC is an alternative c hromatographic tec hnique that offers complementary selectivity to RP and often retains very polar species that cannot
be retained by traditional means. Atlantis HILIC Silica columns were
designed to retain very polar, organic molecules that are too polar
to retain by R P. 1 Unlike the highly aqueous mobile phases required
for polar retention in R P separations, Atlantis HILIC Silica columns
employ highly volatile (>80% organic) mobile phases whic h are ideal
for mass spectrometry (MS) response and sensitivity. Additionally,
direct compatibility with high organic SPE eluates dramatically increases
the number of samples that can be handled, thus substantially increasing
sample throughput.
H2O
H
N
ACN/H2O
(90:10)
H
N
O
Si
NH3
NH3
Si
Si
Si O-
O
H
O
Si
Si
O
O
Si O H
O H
H
O-H
O-
Si
Si
Si
H
O
Si
Si
Si
O
O
Si
Si
O
O
Si
O-
O
Si
H-O
OH-O
Si
Si
Si
Si
O
Si
Si
O
Si O
Si
O
Si
Si
O
O
O
Si
Si
O-
O-
Si
Si
Si
Si
Si
O
O-H
Si
O-H
Si
O
Si
H2O
O-
Acetonitrile/Water (90:10)
H
O
H
H
H
Si
HO H
Si
Water
NH +
O
H-O
O-H
Si
O-
O
N
O-H
Si
Si O
O-
NH +
Si
O
H
N
Si
90% ACN
H
N
Si
OH
H3C
HO
NH
C3H
HO
CH3
NH
1. Bamethan
CH3
2. Albuterol
OH
OH
100
1.84
1
Reversed-Phase
2
2 18
100
0 50% ACN
1.0
2.0
3.0
4.0
5.0 min
1
1.84
2.18
0
1.53
100
2
HILIC
%
95 50% ACN
1.92
0
0.5
136
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0 min
www.waters.com
ANALYTICAL CHROMATOGRAPHY
Atlantis Analytical Columns
Dimension
1.0 x 50 mm
1.0 x 150 mm
2.1 x 10 mm
2.1 x 15 mm
2.1 x 20 mm
2.1 x 20 mm IS
2.1 x 30 mm
2.1 x 50 mm
2.1 x 75 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 20 mm IS
3.0 x 50 mm
3.0 x 75 mm
3.0 x 100 mm
3.0 x 150 mm
3.9 x 20 mm
3.9 x 50 mm
3.9 x 100 mm
3.9 x 150 mm
4.6 x 20 mm
4.6 x 20 mm IS
4.6 x 30 mm
4.6 x 50 mm
4.6 x 75 mm
4.6 x 100 mm
4.6 x 150 mm
1.0 x 50 mm
1.0 x 150 mm
2.1 x 10 mm
2.1 x 15 mm
2.1 x 20 mm
2.1 x 20 mm IS
2.1 x 30 mm
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 20 mm IS
3.0 x 50 mm
3.0 x 100 mm
3.0 x 150 mm
3.0 x 250 mm
3.9 x 20 mm
3.9 x 50 mm
3.9 x 100 mm
3.9 x 150 mm
4.6 x 20 mm
4.6 x 20 mm IS
4.6 x 30 mm
4.6 x 50 mm
4.6 x 75 mm
4.6 x 100 mm
4.6 x 150 mm
4.6 x 250 mm
Type
Particle Size
T3
dC18
HILIC Silica
Column
Column
Guard
Direct Connect
Guard
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Guard
Cartridge
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Column
Column
Guard
Direct Connect
Guard
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Guard
Cartridge
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Column
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
186003713
186003714
1860037561
186003715
186003716
186003717
186005652
186003718
186003719
186003720
186003721
186005653
186003722
186003723
1860037573
1860037583
186003724
186003725
186003726
186003727
186003728
186003729
186003730
186003731
1860037591
186003732
186003733
186003734
186003735
186003736
186003737
186003738
186003739
186003740
186003741
1860037603
1860037613
186003742
186003743
186003744
186003745
186003746
186003747
186003748
186001279
186001283
1860013771
186002064
1860013812
186002058
186001287
186001291
186001295
186001299
186002060
186001389
186001303
186001307
1860013133
1860013854
186001393
186001317
1860013213
186002062
186001325
186001329
186001333
186001337
186001342
186001281
186001285
1860013791
186002065
1860013832
186002059
186001289
186001293
186001297
186001301
186002061
186001391
186001305
186001309
186001311
1860013153
1860013874
186001395
186001319
1860013233
186002063
186001327
186001331
186001335
186001340
186001344
186001346
186002003
1860020051
186002007
186002009
186002011
186002013
186002015
186002017
186002019
1860020213
1860020233
186002025
186002027
186002029
186002031
186002004
1860020061
186002008
186002010
186002012
186002014
186002016
186002018
186002020
1860020223
1860020243
186002026
186002028
186002030
186002032
186002033
Literature
References
Atlantis Columns Brochure,
Literature Reference 720000793EN
Atlantis Columns Applications Notebook,
Literature Reference 720000472EN
Atlantis T3 and ACQUITY UPLC
HSS T3 Columns Toss Sheet,
Literature Reference 720001887EN
Topics in Liquid Chromatography: Part 1.
Designing a Reversed-Phase Column for Polar
Compound Retention W hite Paper,
Literature Reference 720001889EN
Preparative Optimum Bed Density
(OBD) Columns Brochure,
Literature Reference 720002336EN
Interactive Waters Reversed-Phase
Column Selectivity Chart
www.waters.com/selectivitychart
1 Requires
www.waters.com
Particle Size
T3
dC18
HILIC Silica
3 m
5 m
5 m
186003751
186003754
186003755
186002312
186002311
186002313
186002315
186002314
186002316
137
ANALYTICAL CHROMATOGRAPHY
NEW High Strength Silica (HSS) HPLC Columns
Seamless Scalability Across T hree Particle Sizes
Mobile Phase A:
Mobile Phase B:
Injection volumes:
Temperature:
Detection:
Instruments:
Flow Rate
%A
%B
95
5
5
95
5
95
95
5
95
5
0.15
0.10
0.05
0
As large corporations implement UPLC Technology throughout their worldwide organizations there is a transition period during which older HPLC
systems are being phased out and replaced with ACQUITY UPLC systems.
T his transition does not take place overnight, however. During this transition period, c hromatographic methods that are developed using UPLC
Tec hnology must also be compatible with older HPLC systems. To make
this UPLC Tec hnology adoption strategy successful, one must be able to
seamlessly scale between UPLC Technology, HPLC and Preparative LC. T his
is only possible if chromatographic column platforms with HPLC and UPLC
particle sizes exhibit the same selectivities. BEH Technology is the particle
substrate platform tec hnology behind the industry-leading ACQUIT Y
UPLC BEH and XBridge HPLC column families and is a primary reason
behind the success of UPLC Technology. In 2006, Waters commercialized
ACQUIT Y U PLC High Strengt h Silica (HSS) columns based upon t he
1.8 m HSS UPLC particle substrate. Customers who have used and developed methods using ACQUITY UPLC HSS columns require HPLC particle
sizes and columns in order to help with UPLC Technology implementation.
1.7 m, 1.8, m
TRANSFERABILITY
0.5
1.0
1.5
2.0
2.5
3.0
3.5
(mL/min)
0.6
0.6
0.6
0.6
0.6
0.15
0.10
0.05
0
0
10
11
12
24
26
13
Time
(min)
0.00
5.0
6.0
6.01
7.0
0.1% TFA
0.1% TFA in acetonitrile
30 L (4.6 x 150 mm, 5 m)
20 L (4.6 x 100 mm, 3.5 m)
2.1 L (2.1 x 50 mm, 1.8 m)
40 C
UV@230 nm
Alliance 2695 and 2996 PDA (HPLC)
ACQUITY UPLC and PDA (UPLC)
0.15
0.10
0.05
0
0
10
12
14
16
18
20
22
28
30 Min.
Waters developed the HSS HPLC column family in response to customer demands for ACQUITY UPLC HSS
selectivities in HPLC particle and column formats. The result: for a given HSS chemistry, HSS HPLC columns
provide the same selectivity across all particles sizes..
3.5 m, 5 m
2.5, 3.5, 5, 10 m
HSS HPLC columns join the BEH Technology-based XBridge HPLC columns as a
second seamless chromatographic migration path to and from UPLC Technology.
138
www.waters.com
ANALYTICAL CHROMATOGRAPHY
Scale Between Preparative, Analytical and UPLC Separations with HSS Particles
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
Paroxetine
0.005
0.000
4.0
4.5
2
5.0
5.5
6.0
6.5
6.9 min
0.010
AU
UPLC System:
0.005
Paroxetine
Detection:
HPLC System:
0.1% TFA
0.1% TFA in acetonitrile
30 C
50:50 MeOH:H2O
Related Compound
1: 0.002 mg/mL
Paroxetine; 0.2 mg/mL
Related Compound
2: 0.002 mg/mL
UV@295 nm
Alliance 2695
with 2996 PDA
ACQUITY UPLC
with PDA
AU
Mobile Phase A:
Mobile Phase B:
Temperature:
Sample Diluent:
Concentrations:
0.000
0
10
12
14
16
18
20
20.8 min
0.005
0.000
0
Paroxetine
AU
0.010
9.7 min
UPLC Separation
0.005
0.000
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
Paroxetine
AU
0.010
1.8
2.0
2.2
2.4
2.5 min
Type
2-pk1
Guard Cartridge
Column
Column
Column
Column
Column
Guard Cartridge 2-pk1
Column
Column
Column
Column
Column
Guard Cartridge 2-pk1
Column
Column
Column
Particle Size
HSS C18
HSS C18 SB
HSS T3
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
5 m
186004764
186004765
186004766
186005642
186004762
186004763
186004769
186004771
186004772
186004767
186004768
186004770
186004774
186004852
186004773
186004775
186004745
186004746
186004747
186005643
186004743
186004744
186004750
186004752
186004753
186004748
186004749
186004751
186004755
186004757
186004754
186004756
186004782
186004783
186004784
186005641
186004780
186004781
186004787
186004789
186004790
186004785
186004786
186004788
186004792
186004794
186004791
186004793
Requires 3.0 x 20 mm/4.6 x 20 mm Universal Sentry Guard Holder, Part No. WAT046910
Literature
References
High Strength Silica (HSS)
HPLC Columns Toss Sheet,
Literature Reference 720003241EN
ACQUITY UPLC Column Strategies Brochure,
Literature Reference 720003271EN
Interactive Waters Reversed-Phase
Column Selectivity Chart,
www.waters.com/selectivitychart
Type
Guard Cartridge
Prep Column
Prep Column
Prep Column
2-pk2
Particle Size
HSS C18
HSS C18 SB
HSS T3
5 m
5 m
5 m
5 m
186004776
186004778
186004779
186004777
186004758
186004760
186004761
186004759
186004795
186004797
186004798
186004796
www.waters.com
139
aNaLYTICaL ChROMaTOGRaPhY
XTerra Columns
T he Most Successful Column in Over 30 Years
of Stationary-Phase Technology
T he universal acceptance by pharmaceutical scientists in drug discovery,
method development, and isolation and purification has enabled XTerra
columns to become the fastest selling and most successful column product
in the world of chromatography. T he reason: the enabling science of Hybrid
Particle Tec hnology. Drug discovery scientists benefit from the DMSO
injection-resistance of the rugged XTerra particle. Method development
scientists are able to operate at whatever pH is necessary for optimal
selectivity due to XTerra columns wide pH range. Isolation and purification
scientists can load up to 60x more material per injection because of the
high mass loading capacity of XTerra columns.
Physical Characteristics
%C
10
10
10
Flow Rate:
Injection Volume:
Temperature:
Detection:
Instrument:
1.0 mL/min
15 mL
30 C
UV @ 254nm
Waters Alliance 2695,
996 PDA
Peaks:
1. Suprofen
2. Tolmetin
3. Naproxen
4. Fenoprofen
5. Ibuprofen
6. Diclofenac
Packing
Particle
Shape
Particle
Size(s)
Pore
Size
Carbon
Load
Endcapped
XTerra MS C18
XTerra MS C8
Spherical
Spherical
2.5, 3.5, 5, 10 m
2.5, 3.5, 5, 10 m
125
125
15.5%
12.0%
yes
yes
XTerra RP18
XTerra RP8
Spherical
Spherical
3.5, 5, 10 m
3.5, 5, 10 m
125
125
15.0%
13.5%
yes
yes
XTerra Phenyl
Spherical
3.5, 5 m
125
12.0%
yes
XTerra MS C8
O
Si
XTerra RP8
CH3
O
Si
CH3
XTerra Phenyl
Si
CH3
O
O
CH2 CH
CH3
10
15
20
25 Min.
Patent # 6,686,035 B2
The column stationary phase chemistries of XTerra columns are key
components in an overall method development plan.
140
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
Ordering Information
XTerra Capillary and Narrowbore Columns
Dimension
Type
75 m x 50 mm
75 m x 100 mm
75 m x 150 mm
100 m x 50 mm
100 m x 100 mm
100 m x 150 mm
150 m x 50 mm
150 m x 100 mm
150 m x 150 mm
300 m x 50 mm
300 m x 100 mm
300 m x 150 mm
300 m x 50 mm
300 m x 100 mm
300 m x 150 mm
1.0 mm x 50 mm
1.0 mm x 50 mm
1.0 mm x 100 mm
1.0 mm x 150 mm
2.1 mm x 10 mm
2.1 mm x 10 mm
2.1 mm x 15 mm
2.1 mm x 15 mm
2.1 mm x 15 mm
2.1 mm x 15 mm
2.1 mm x 20 mm
2.1 mm x 20 mm
2.1 mm x 20 mm
2.1 mm x 20 mm
2.1 mm x 20 mm
2.1 mm x 30 mm
2.1 mm x 30 mm
2.1 mm x 50 mm
2.1 mm x 50 mm
2.1 mm x 50 mm
2.1 mm x 50 mm
2.1 mm x 50 mm
2.1 mm x 100 mm
2.1 mm x 100 mm
2.1 mm x 100 mm
2.1 mm x 100 mm
2.1 mm x 150 mm
2.1 mm x 150 mm
2.1 mm x 150 mm
2.1 mm x 150 mm
2.1 mm x 250 mm
2.1 mm x 250 mm
Capillary
Capillary
Capillary
Capillary
Capillary
Capillary
Capillary
Capillary
Capillary
Capillary
Capillary
Capillary
Capillary
Capillary
Capillary
Column
Column
Column
Column
Guard
Guard
Direct Connect Column
Direct Connect Column
Direct Connect Column
Direct Connect Column
IS Column
IS Column
Guard
IS Column
Guard
Column
Column
Column
Column
Cartridge
Column
Cartridge
Column
Cartridge
Column
Cartridge
Column
Cartridge
Column
Cartridge
Column
Cartridge
Particle Size
MS C18
MS C8
RP18
RP8
Phenyl
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
2.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
2.5 m
3.5 m
5 m
10 m
2.5 m
3.5 m
3.5 m
5 m
5 m
2.5 m
3.5 m
2.5 m
3.5 m
3.5 m
5 m
5 m
3.5 m
3.5 m
5 m
5 m
3.5 m
3.5 m
5 m
5 m
5 m
5 m
186002198
186002199
186002200
186002210
186002211
186002212
186002469
186002470
186002471
186002599
186002600
186002601
186002602
186002603
186002604
186000979
186000386
186000390
186000394
1860006321
1860006481
186000900
186001908
186001907
186001906
186001921
186001923
1860006362
186001979
1860006522
186000592
186000398
186000594
186000400
1860004983
186000446
1860005383
186000404
1860005023
186000450
1860005423
186000408
1860005063
186000454
1860005463
186000458
1860005503
186000387
186000391
186000395
1860006331
1860006491
186001922
186001924
1860006372
186001980
1860006532
186000593
186000399
186000595
186000401
1860004993
186000447
1860005393
186000405
1860005033
186000451
1860005433
186000409
1860005073
186000455
1860005473
186000459
1860005513
186000388
186000392
186000396
1860006341
1860006501
186001925
1860006382
186001982
1860006542
186000402
1860005003
186000448
1860005403
186000406
1860005043
186000452
1860005443
186000410
1860005083
186000456
1860005483
186000460
1860005523
186000389
186000393
186000397
1860006351
1860006511
186001926
1860006392
186001983
1860006552
186000403
1860005013
186000449
1860005413
186000407
1860005053
186000453
1860005453
186000411
1860005093
186000457
1860005493
186000461
1860005533
186001188
186001192
186001189
186001193
186001179
186001185
186001180
186001186
186001181
186001187
Also available in Preparative Optimum Bed Density (OBD) Columns, See Page 189
www.waters.com
141
aNaLYTICaL ChROMaTOGRaPhY
XTerra Analytical Columns
Dimension
Type
3.0 x 20 mm
3.0 x 20 mm
3.0 x 20 mm
3.0 x 20 mm
3.0 x 20 mm
3.0 x 20 mm
3.0 x 30 mm
3.0 x 30 mm
3.0 x 50 mm
3.0 x 50 mm
3.0 x 50 mm
3.0 x 50 mm
3.0 x 50 mm
3.0 x 100 mm
3.0 x 100 mm
3.0 x 100 mm
3.0 x 100 mm
3.0 x 150 mm
3.0 x 150 mm
3.0 x 150 mm
3.0 x 150 mm
3.0 x 250 mm
3.0 x 250 mm
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
3.9 x 50 mm
3.9 x 50 mm
3.9 x 100 mm
3.9 x 100 mm
3.9 x 100 mm
3.9 x 150 mm
3.9 x 150 mm
3.9 x 150 mm
4.6 x 10 mm
4.6 x 10 mm
4.6 x 20 mm
4.6 x 20 mm
4.6 x 20 mm
4.6 x 20 mm
4.6 x 30 mm
4.6 x 30 mm
4.6 x 30 mm
4.6 x 50 mm
4.6 x 50 mm
4.6 x 50 mm
4.6 x 50 mm
4.6 x 50 mm
4.6x 100 mm
4.6 x 100 mm
4.6 x 100 mm
4.6 x 150 mm
4.6 x 150 mm
4.6 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
4.6 x 250 mm
4.6 x 250 mm
IS Column
Cartridge
IS Column
Guard
IS Column
Guard
Column
Column
Column
Column
Cartridge
Column
Cartridge
Column
Cartridge
Column
Cartridge
Column
Cartridge
Column
Cartridge
Column
Cartridge
IS Column
IS Column
Guard
IS Column
Guard
Cartridge
Cartridge
Column
Cartridge
Column
Column
Column
Cartridge
Guard
Guard
IS Column
Cartridge
IS Column
IS Column
Column
Column
Column
Column
Column
Cartridge
Column
Cartridge
Column
Cartridge
Column
Column
Cartridge
Column
Cartridge
Column
Column
Cartridge
4
5
Particle Size
MS C18
MS C8
RP18
RP8
Phenyl
2.5 m
2.5 m
3.5 m
3.5 m
5 m
5 m
2.5 m
3.5 m
2.5 m
3.5 m
3.5 m
5 m
5 m
3.5 m
3.5 m
5 m
5 m
3.5 m
3.5 m
5 m
5 m
5 m
5 m
2.5 m
3.5 m
3.5 m
5 m
5 m
3.5 m
5 m
3.5 m
3.5 m
5 m
3.5 m
5 m
5 m
3.5 m
5 m
2.5 m
2.5 m
3.5 m
5 m
2.5 m
3.5 m
5 m
2.5 m
3.5 m
3.5 m
5 m
5 m
3.5 m
3.5 m
5 m
3.5 m
3.5 m
5 m
5 m
3.5 m
5 m
5 m
186001972
1860005884
186001974
1860006404
186001984
1860006564
186000596
186000412
186000598
186000414
1860005105
186000462
1860005545
186000418
1860005145
186000466
1860005585
186000422
1860005185
186000470
1860005625
186000474
1860005665
186001899
186001900
1860006444
186001988
1860006604
1860008175
1860008155
186000426
1860005225
186000478
1860005705
1860019276
1860019206
186001889
1860005904
186001891
186001992
186000600
186000430
186000878
186000602
186000432
1860005265
186000482
1860005745
186000436
1860005305
186000486
186000440
1860005345
186000490
1860005785
186001470
186000494
1860005825
186001973
1860005894
186001975
1860006414
186001985
1860006574
186000597
186000413
186000599
186000415
1860005115
186000463
1860005555
186000419
1860005155
186000467
1860005595
186000423
1860005195
186000471
1860005635
186000475
1860005675
186001897
186001898
1860006454
186001989
1860006614
1860008185
1860008165
186000427
1860005235
186000479
1860005715
1860014346
186001890
1860005914
186001892
186001993
186000601
186000431
186000879
186000603
186000433
1860005275
186000483
1860005755
186000437
1860005315
186000487
186000441
1860005355
186000491
1860005795
186001471
186000495
1860005835
186001976
1860006424
186001986
1860006584
186000416
1860005125
186000464
1860005565
186000420
1860005165
186000468
1860005605
186000424
1860005205
186000472
1860005645
186000476
1860005685
186001902
1860006464
186001990
1860006624
186000428
1860005245
186000480
1860005725
186001893
186001994
186001910
186001909
186000434
1860005285
186000484
1860005765
186000438
1860005325
186000488
186000442
1860005365
186000492
1860005805
186001472
186000496
1860005845
186001977
1860006434
186001987
1860006594
186000417
1860005135
186000465
1860005575
186000421
1860005175
186000469
1860005615
186000425
1860005215
186000473
1860005655
186000477
1860005695
186001901
1860006474
186001991
1860006634
186000429
1860005255
186000481
1860005735
186001894
186001995
186001912
186001911
186000435
1860005295
186000485
1860005775
186000439
1860005335
186000489
186000443
1860005375
186000493
1860005815
186001473
186000497
1860005855
186001190
186001194
186001141
186001148
186001142
186001149
186001143
186001150
186001151
186001191
186001195
186001204
186001203
186001177
186001183
186001178
186001184
186001144
186001139
186001145
186001140
186001146
186001474
186001147
Requires Part No. WAT046910 (universal, use with standard columns) or Part No. WAT046905 (integrated into cartridge column)
Requires End Connector Kit, Part No. WAT037525 6 Requires Inline Guard Cartridge Holder, Part No. PSS830008
142
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
XTerra Oligonucleotide Purification Columns
Dimension
4.6 x 50 mm
10 x 50 mm
Type
Particle Size
MS C18
Column
Column
2.5 m
2.5 m
186000602
186000982
Type
Particle Size
MS C18
MS C8
RP18
RP8
Phenyl
MV Kit
MV Kit
MV Kit
MV Kit
MV Kit
MV Kit
MV Kit
3.5 m
5 m
5 m
3.5 m
3.5 m
5 m
5 m
Custom
186000827
186000828
Custom
186000826
186000829
186000830
186000831
Custom
Custom
186000832
Custom
Custom
186000833
Custom
Custom
Custom
Custom
186000861
186000862
186000863
186000834
Custom
186000836
186000835
Custom
Custom
Custom
Custom
Custom
Custom
Custom
186002234
186002235
186002236
Also available in Preparative Optimum Bed Density (OBD) Columns, See Page 189
Literature
References
XTerra Columns Brochure,
Literature Reference 720000424EN
XTerra Applications Notebook,
Literature Reference 720000502EN
www.waters.com
Description
Part No.
715001531
715002531
143
aNaLYTICaL ChROMaTOGRaPhY
Symmetry Columns
Setting the Standard for Reproducibility
As todays c hemists establish new analytical methods for the latest
pharmaceutical and biopharmaceutical products, the selection of a
reproducible HPLC column is essential. T he selected column needs to
provide the same c hromatographic results over the life of the new
drug product. Symmetry columns demonstrate superior reproducibility
over many years. Batc hes randomly selected over a five year period
demonstrate excellent reproducibility in the example shown.
Reproducibility is our number one priority in supplying Symmetry columns.
T his excellent reproducibility is a result of our commitment to maintaining the tightest specifications in the HPLC column industry. Symmetry
columns are engineered with high purity raw materials, tightly controlled
manufacturing processes and column packing procedures that provide
todays scientists with the best, most reproducible HPLC column available.
Symmetry C18,
4.6 x 150 mm, 5 m
WAT045905
H2O
MeCN
pH 3.75; 100 mM
NH4COOH in H2O
1.4 mL/min
30% A; 60% B; 10% C
5.0 L
30 C
UV @ 233 nm
Waters Alliance HT 2795
Waters 996 PDA detector
Sampling Rate: 5 pts./sec
Part No.:
Mobile Phase A:
Mobile Phase B:
Mobile Phase C:
Flow Rate:
Isocratic:
Injection Volume:
Temperature:
Detection:
Instrumentation:
0.7%
0.8%
0.6%
0.7%
2
3
Our goal is to supply a family of HPLC columns that you can rely on for
rugged and robust methods. Symmetry columns let you increase your
laboratorys productivity, and allow easier method transfer between labs
and around the globe.
Physical Characteristics
Chemistry
Particle
Size
Particle
Shape
Pore
Size
Carbon
Load
Endcapped
Symmetry
C18
C8
3.5, 5 m
3.5, 5 m
Spherical
Spherical
100
100
19%
12%
Yes
Yes
SymmetryPrep
C18
C8
7 m
7 m
Spherical
Spherical
100
100
19%
12%
Yes
Yes
3.5, 5, 7 m Spherical
3.5, 5, 7 m Spherical
100
100
17%
15%
Yes
Yes
300
300
8.5%
2.8%
Yes
Yes
Packing
SymmetryShield
RP18
RP8
Symmetry300
C18
C4
144
3.5, 5 m
3.5, 5 m
Spherical
Spherical
1.5
3.0
4.5
6.0
7.5
9.0
10.5
12.0
13.5
15.0 Min.
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aNaLYTICaL ChROMaTOGRaPhY
T he Benefit of Narrow Specification Ranges
Symmetry columns have the narrowest ligand surface coverage specifications in the industry. Surface coverage (ligand density) is one of the most
important parameters affecting reproducibility. T he impact of a small
variation in surface coverage on the amitriptyline/acenaphthene alpha
specification for Symmetry C 18 columns is shown here. Narrow column
specifications benefit the c hromatographer in ac hieving reproducible
results. T he surface coverage requirements of Symmetry are the tightest
specifications in the industry, resulting in minimal shifts in resolution
and more reproducible batch-to-batch and column-to-column results.
Symmetry C18
3.31
3.75
2.75
Column X
2.6
no upper limit
Column Y
1.5
2.0
2.5
3.0
3.5
4.0
2
1.8
4.14
Symmetry C18
1.6
4.14
2.86
1.4
4.14
2.86
1.2
Specification Ranges
for Symmetry C18
Specification X
0.8
1.5
2.0
2.5
3.0
3.5
4.0
2.86
N
4
)( a-1a )( KK+1 )
2
Specification Y
R = resolution
N = number of peaks
a = alpha value
k = peak retention
4.70
0.81
4.70
0.81
4.70
0.81
5.37
5.37
5.37
Maximum Resolution
Minimum Resolution
Long Lifetime
Columns that last longer save you money. Symmetry columns can last over
10,000 injections with minimal loss in efficiency, minimal increase in
backpressure and minimal change in retention time.
Column:
Mobile Phase:
Flow Rate:
Injection Volume:
Temperature:
Detector:
Analytes:
Injection 10,000
Start
0
www.waters.com
10 Min.
The long-term mechanical stability of Symmetry columns is demonstrated in this analysis of sulfa drugs.
Over 10,000 injections were run with minimal loss in column efficiency. The use of the Sentry guard
column is critical to achieving these types of column lifetimes.
145
aNaLYTICaL ChROMaTOGRaPhY
Symmetry Columns and Cartridges
Symmetry Analytical Columns
Dimension
Symmetry IS Columns
Particle Size
C18
C8
1.0 x 50 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 50 mm
3.0 x 100 mm
3.0 x 150 mm
4.6 x 30 mm
4.6 x 50 mm
4.6 x 75 mm
4.6 x 100 mm
4.6 x 150 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
WAT106056
WAT248059
WAT058973
WAT200650
WAT058965
WAT106005
186002612
186000696
186000695
186000271
WAT200625
WAT066224
WAT066220
WAT200632
WAT106052
WAT248072
WAT058977
WAT200624
WAT058961
WAT106011
186002613
186000698
186000697
186000270
WAT200620
WAT066200
WAT066204
WAT200630
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 150 mm
3.0 x 250 mm
3.9 x 150 mm
4.6 x 50 mm
4.6 x 100 mm
4.6 x 150 mm
4.6 x 250 mm
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
186000206
186002608
WAT056975
WAT054200
186000690
WAT046980
186000207
186002616
WAT045905
WAT054275
186000212
186002609
WAT056955
WAT054230
186000691
WAT046970
186000213
186002617
WAT045995
WAT054270
Dimension
Particle Size
C18
C8
2.1 x 20 mm
3.0 x 20 mm
3.9 x 20 mm
4.6 x 20 mm
3.5 m
3.5 m
3.5 m
3.5 m
186002066
186002074
186002082
186002090
186002067
186002075
186002083
186002091
2.1 x 20 mm
3.0 x 20 mm
3.9 x 20 mm
4.6 x 20 mm
5 m
5 m
5 m
5 m
186002070
186002078
186002086
186002094
186002071
186002079
186002087
186002095
Particle Size
C18
C8
3.5 m
5 m
WAT106127
WAT054225
WAT106128
WAT054250
2.1 x 10 mm
3.9 x 20 mm
Particle Size
C18
C8
2.1 x 20 mm
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
4.6 x 75 mm
4.6 x 100 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
186000269
186000152
186000151
186000150
WAT066260
WAT066265
186000268
186000149
186000153
186000148
WAT066210
WAT066215
3.9 x 50 mm
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
5 m
5 m
5 m
5 m
WAT054220
WAT054205
WAT054210
WAT054215
WAT054240
WAT054235
WAT054255
WAT054245
Type
Particle Size
C18
C8
4.6 x 150 mm
2.1 x 150 mm
3.0 x 150 mm
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
Column
Column
Column
Column
Column
Column
3.5 m
5 m
5 m
5 m
5 m
5 m
WAT094240
WAT094234
WAT054446
WAT047210
WAT054448
WAT054450
WAT094237
WAT094231
WAT054434
WAT046955
WAT054435
WAT054438
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
Cartridge*
Cartridge*
Cartridge*
5 m
5 m
5 m
WAT054452
WAT054454
WAT054456
WAT054440
WAT054442
WAT054444
Literature
References
Symmetry Family of Columns Brochure,
Literature Reference 720000454EN
Symmetry Family of Columns
Applications Notebook,
Literature Reference 720000593EN
Interactive Waters Reversed-Phase
Column Selectivity Chart,
www.waters.com/selectivitychart
146
www.waters.com
ANALYTICAL CHROMATOGRAPHY
SymmetryShield Columns
Excellent Peak Shape
SymmetryShield RP18
Brand L
1.66
Brand HE
1.75
Brand DO
1.78
Brand DA
1.9
Polar
Group
Si
1.5
Brand HP
Shield Technology
1.2
1.95
Brand I
2.39
Brand P
3.6
Brand E
CH3
CH3
a: SymmetryShield RP18
1
Flow Rate:
Detection:
Temperature:
Analytes:
6
7
10
b: Symmetry C18
SymmetryShield columns with Shield Technology are the perfect choice for
applications that require low organic, highly aqueous mobile-phase conditions. T hese reversed-phase particles are water-wettable allowing better
interaction between the particles and your sample. T he result is stable
retention in highly aqueous mobile phases with superior peak shape.
www.waters.com
15
20
25
30 Min.
2
3
AU
a. SymmetryShield RP18
3.9 x 150 mm
b. Leading Brand C18
65% methanol; 35% 20 mM
KH2PO4/K2HPO4 at pH 7
1.0 mL/min
UV @ 254 nm
23 C
1. Uracil
2. Propranolol
3. Butylparaben
4. Dipropylphthalate
5. Naphthalene
6. Amitriptyline
7. Acenaphthene
Mobile Phase:
Unique Selectivity
Column:
AU
2
USP Tailing Factor
10
15
20
25
30 Min.
147
ANALYTICAL CHROMATOGRAPHY
SymmetryShield ColumnsNo Pore Dewetting in Highly Aqueous Mobile Phases
SymmetryShield RP8 ColumnsNo Pore Dewetting!
Column:
Amoxicillin
10 Min.
Column:
Amoxicillin
Initial
Initial
No change in
retention time
0
SymmetryShield RP8
4.6 x 150 mm, 5 m
10 Min.
SymmetryShield Columns
Dimension
SymmetryShield IS Columns
Particle Size
RP18
RP8
1.0 x 50 mm
1.0 x 150 mm
2.1 x 30 mm
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 50 mm
3.0 x 100 mm
3.0 x 150 mm
4.6 x 50 mm
4.6 x 75 mm
4.6 x 100 mm
4.6 x 150 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
186000175
186000176
186000171
186000172
186000173
186000174
186002614
186000700
186000699
186000177
186000178
186000179
186000180
WAT106060
WAT106048
WAT106001
WAT094257
WAT058969
WAT106008
186002615
186000703
186000702
WAT094260
WAT094263
WAT094266
WAT094269
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
3.0 x 150 mm
3.0 x 250 mm
3.9 x 150 mm
4.6 x 50 mm
4.6 x 100 mm
4.6 x 150 mm
4.6 x 250 mm
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
186000217
186000998
186000111
186000692
186000693
186000108
186000218
186002618
186000109
186000112
186000223
186002611
WAT094245
WAT094243
186000694
WAT200655
186000224
186002619
WAT200662
WAT200670
Dimension
Particle Size
RP18
RP8
2.1 x 20 mm
3.0 x 20 mm
3.9 x 20 mm
4.6 x 20 mm
3.5 m
3.5 m
3.5 m
3.5 m
186002068
186002076
186002084
186002092
186002069
186002077
186002085
186002093
2.1 x 20 mm
3.0 x 20 mm
3.9 x 20 mm
4.6 x 20 mm
5 m
5 m
5 m
5 m
186002072
186002080
186002088
186002096
186002073
186002081
186002089
186002097
Particle Size
RP18
RP8
3.5 m
3.5 m
5 m
186000169
186000701
186000107
WAT106129
186000704
WAT200675
2.1 x 10 mm
3.9 x 20 mm
3.9 x 20 mm
Dimension
Particle Size
RP18
RP8
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
4.6 x 75 mm
4.6 x 100 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
186000168
186000167
186000166
186000183
186000170
186000147
186000146
186000145
WAT094272
WAT094275
3.9 x 50 mm
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
5 m
5 m
5 m
5 m
186000106
186000110
186000113
WAT094248
WAT200658
WAT200665
WAT200661
148
Type
Particle Size
RP18
RP8
2.1 x 150 mm
4.6 x 150 mm
2.1 x 150 mm
3.0 x 150 mm
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
Column
Column
Column
Column
Column
Column
Column
3.5 m
3.5 m
5 m
5 m
5 m
5 m
5 m
186000182
186000181
186000100
186000104
186000103
186000102
WAT094278
WAT094254
WAT094251
WAT210594
WAT210588
WAT210591
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
Cartridge*
Cartridge*
Cartridge*
5 m
5 m
5 m
186000105
186000101
186000114
WAT210582
WAT210585
WAT210579
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aNaLYTICaL ChROMaTOGRaPhY
Symmetry300 Columns
High Recoveries of Proteins and Peptides
Symmetry300 columns are wide-pore, reversed-phase columns whic h
are built on t he Symmetry column platform, t he standard t hat t he
pharmaceutical industry has come to rely on for sensitive, rugged and
robust HPLC analyses.
Peptide Separation at High Mass Load (0.1% TFA)
Columns:
Mobile Phase A:
Mobile Phase B:
Flow Rate:
Gradient:
4.6 x 50 mm, 5 m
0.1% TFA in H2O
0.08% TFA in ACN
0.75 ml/min
Time
Profile
(min)
%A
%B
0.00
100
0
25.00 50
50
Temperature:
Detection:
Instrument:
Symmetry300 C18,
4.6 x 50 mm column
P = 98.8*
2
1
40 C
UV 220 nm
Waters Alliance 2690
7
4
5 6
10
15
20 Min.
MW
986.6
1001.5
1060.2
1046.2
1296.5
1347.6
1759.0
3495.9
2845.8
pI
12.04
9.34
12.00
7.35
7.51
11.20
7.61
7.51
12.06
Hydrophobicity Number of
(Meek)
Amino Acids
2.8
9
3.0
10
42.6
9
43.7
8
54.0
10
52.7
11
63.5
14
129.4
30
92.3
26
pI = Isoelectric pH
Symmetry300 C18 columns provide better peak shape and peak capacity
compared to other leading brands when separating biologically active
peptides. In the examples shown, Symmetry300 columns demonstrate
superior performance when simultaneously separating hydrophilic and
hydrophobic peptides, as well as neutral and highly basic peptides. Also,
hydrophobicity (Meek retention prediction 1) is included. T he lower
hydrophobicity values indicate the peptide is more hydrophilic.
4.6 x 50 mm, 5 m
0.1% formic acid in H2O
0.08% formic acid in ACN
0.75 ml/min
Gradient:
Injection Volume:
Temperature:
Detection:
Instrument:
6
3
Time
Profile
(min)
%A
%B
0
100
0
25
50
50
20 L
40 C
UV 220 nm
Waters Alliance 2690
Symmetry300 C18 ,
4.6 x 50 mm
P = 93.1*
T he heart of the column is the high purity base deactivated silica. T he silica
used in Symmetry300 columns is synthesized using ultrapure organic
reagents, resulting in high purity with very low silanol activity.
In addition, when combined with the high surface coverage of the bonded
phase the result is higher recoveries of proteins and peptides even when
injected at low concentrations.
Symmetry300 columns deliver the sharpest peaks for biomolecules and
deliver more peak capacity for complex tryptic maps. T hey are packed to
the highest efficiency and reproducibility of any product on the market.
T his results in improved precision for impurity and stability assays, more
accurate quantitation, and adaptability to more rugged methods.
Unsurpassed Reproducibility
Cytochrome c Tryptic Map Q.C. Test
Column:
Part Number:
Mobile Phase A:
Mobile Phase B:
Flow Rate:
Gradient:
Symmetry300 C18
3.9 x 150 mm, 5 m
WAT106154
0.05% TFA in water
0.1% TFA in acetonitrile
1.0 mL/min
Time
Profile
(min)
%A
%B
0
100
0
45
72
28
20 L
35 C
UV @ 220 nm
Injection Volume:
Temperature:
Detection:
Batch
Batch107
107
Batch
106
Batch
106
10
15
Batch
104
Batch
104
20 Min.
Batch
103
Batch
103
* Note: Peak Capacity (P) is defined as the number of theoretical peaks that can
fit into a chromatographic run within a certain gradient time.
10
30
50 Min.
P = 1 + (tg/w5s) Where tg = gradient time w5s = peak measured at 4.4% of peak height
www.waters.com
149
aNaLYTICaL ChROMaTOGRaPhY
Excellent Protein Separations
Columns:
Mobile Phase A:
Mobile Phase B:
Flow Rate:
Gradient:
4.6 x 50 mm, 5 m
0.1% TFA in water
0.08% TFA in acetonitrile
0.75 mL/min
Time
Profile
(min)
%A
%B
0.0
80
20
15.0
35
65
Injection Volume:
Temperature:
Detection:
Instrument:
Analytes:
20 L
40 C
UV @ 220 nm
Waters Alliance 2790, 996 PDA
1. RNAse
2. BSA
3. B-Lactoglobulin
4. Ovalbumin
Symmetry300 C4
P = 138.56
Particle Size
C18
C4
1.0 x 150 mm
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
4.6 x 50 mm
4.6 x 75 mm
4.6 x 100 mm
4.6 x 150 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
186000185
186000187
186000188
186000200
186000201
186000189
186000190
186000197
186000276
186000277
186000278
186000279
186000280
186000281
186000282
186000283
2.1 x 150 mm
3.9 x 150 mm
4.6 x 50 mm
4.6 x 150 mm
4.6 x 250 mm
5 m
5 m
5 m
5 m
5 m
WAT106172
WAT106154
WAT106209
WAT106157
WAT106151
186000285
186000286
186000287
186000288
186000289
Particle Size
C18
2.1 x 50 mm
4.6 x 75 mm
3.5 m
3.5 m
186000199
186000192
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
5 m
5 m
5 m
WAT106169
WAT106163
WAT106160
150
10
12
14 Min.
Particle Size
C18
C4
3.5 m
5 m
186000198
WAT106166
186000275
186000284
2.1 x 10 mm
3.9 x 20 mm
Type
Particle Size
C18
C4
2.1 x 150 mm
4.6 x 150 mm
Column
Column
3.5 m
3.5 m
186000194
186000195
186000290
186000291
2.1 x 150 mm
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
Column
Column
Column
Column
5 m
5 m
5 m
5 m
WAT106193
WAT106187
WAT106190
WAT106184
186000292
186000293
186000294
186000295
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
Cartridge*
Cartridge*
Cartridge*
5 m
5 m
5 m
WAT106181
WAT106175
WAT106178
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
Symmetry NanoEase Nano and Capillary Columns
NanoEase columns contain high performance, small particle sorbents.
Exact manufacturing and QC production procedures ensure consistent
column performance. Superior column longevity is ensured by a novel
frit tec hnology that minimizes bed disturbance throughout the life of
the column.
Description
Gradient:
Flow Rate:
System:
5-50% B in 60 min
400 nL/min
Waters CapLC operated in
split flow mode, Q-Tof micro
100
TOF MS ES+
TIC
9.78e4
Inj #4
100
%
Inj #3
TOF MS ES+
TIC
1.48e5
Inj #2
TOF MS ES+
TIC
1.71e5
100
%
0
100
TOF MS ES+
TIC
1.69e5
Inj #1
%
0
10
www.waters.com
20
30
40
50
60 Min.
Column i.d.
Length
Particle Size
Part No.
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
75 m
75 m
75 m
75 m
75 m
75 m
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
186002188
186002189
186002190
186002191
186002192
186002193
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry300 C18
100 m
100 m
100 m
100 m
50 mm
100 mm
150 mm
150 mm
3.5 m
3.5 m
3.5 m
3.5 m
186002201
186002202
186002203
186002206
Symmetry C18
Symmetry C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
150 m
150 m
150 m
150 m
150 m
50 mm
150 mm
50 mm
100 mm
150 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
186002459
186002461
186002462
186002463
186002464
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
186002581
186002582
186002583
186002584
186002585
186002586
186002587
186002588
186002589
186002590
186002591
186002592
151
aNaLYTICaL ChROMaTOGRaPhY
Physical Characteristics
Particle
Shape
Surface
Area
Pore
Size
Pore
Volume
Particle
Size
Carbon
Load
Ligand
Coverage
Endcapped
Spherical
Spherical
Spherical
220 m2/g
220 m2/g
220 m2/g
80
80
80
0.50 ml/g
0.50 ml/g
0.50 ml/g
3, 5, 10 m
3, 5, 10 m
5 m
11.5%
6.2%
11.5%
2.98 Mol/m2
1.49 Mol/m2
2.98 Mol/m2
yes
no
yes
Waters Spherisorb C8
Waters Spherisorb C6
Waters Spherisorb C1
Spherical
Spherical
Spherical
220 m2/g
220 m2/g
220 m2/g
80
80
80
0.50 ml/g
0.50 ml/g
0.50 ml/g
3, 5, 10 m
3, 5, 10 m
3, 5, 10 m
5.75%
4.7%
2.15%
3.12 Mol/m2
3.36 Mol/m2
2.97 Mol/m2
yes
yes
no
Spherical
Spherical
Spherical
220 m2/g
220 m2/g
220 m2/g
80
80
80
0.50 ml/g
0.50 ml/g
0.50 ml/g
3, 5, 10 m
3, 5, 10 m
3, 5, 10 m
1.9%
2.5%
3.1%
2.64 Mol/m2
1.72 Mol/m2
3.29 Mol/m2
no
no
no
Spherical
Spherical
220 m2/g
220 m2/g
80
80
0.50 ml/g
0.50 ml/g
5 m
3, 5, 10 m
5%
1.15 Mol/m2
yes
Spherical
Spherical
220 m2/g
220 m2/g
80
80
0.50 ml/g
0.50 ml/g
5, 10 m
5, 10 m
4%
4%
no
no
Product Description
Waters Spherisorb Phases
152
Analytical Chromatography
4.6 mm
x 100 mm
4.6 mm
x 150 mm
4.6 mm
x 200 mm
4.6 mm
x 250 mm
PSS830712
PSS833612
PSS833512
PSS830212
PSS832513
PSS830713
PSS832813
PSS833213
PSS833013
PSS833613
PSS833813
PSS833513
PSS830213
PSS832514
PSS830714
PSS832514
PSS830214
PSS832515
PSS830715
PSS832815
PSS833215
PSS833015
PSS833615
PSS833815
PSS833515
PSS830215
PSS833912
PSS833913
PSS837613
PSS833915
PSS837615
Ion Exchange
S10 SAX
S10 SCX
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aNaLYTICaL ChROMaTOGRaPhY
Waters Spherisorb 5 m Columns and Cartridges
Columns
Chemistry
S5 ODS2
S5 ODS1
S5 ODS B
S5 C8
S5 C6
S5 C1
S5 NH2
S5 P (Phenyl)
S5 CN Normal Phase
S5 CN Reversed Phase
S5 W (Silica)
Cartridge Columns
4.0 mm
x 125 mm
4.0 mm
x 250 mm
4.6 mm
x 50 mm
4.6 mm
x 100 mm
4.6 mm
x 150 mm
4.6 mm
x 250 mm
4.6 mm
x 50 mm
4.6 mm
x 100 mm
4.6 mm
x 150 mm
4.6 mm
x 250 mm
PSS845543
PSS845541
PSS845280
PSS845284
PSS845288
PSS845300
PSS845292
PSS845296
PSS845539
PSS845277
PSS845542
PSS845281
PSS845285
PSS845289
PSS845301
PSS845293
PSS845297
PSS845540
PSS831911
PSS830611
PSS831811
PSS831011
PSS832611
PSS831111
PSS830911
PSS830111
PSS831912
PSS830612
PSS831812
PSS831012
PSS832612
PSS831112
PSS830812
PSS830912
PSS830112
PSS831913
PSS830613
PSS831813
PSS831013
PSS832613
PSS831113
PSS830813
PSS830913
PSS830908
PSS830113
PSS831915
PSS830615
PSS831815
PSS831015
PSS832615
PSS831115
PSS830815
PSS830915
PSS830909
PSS830115
PSS839536
PSS839506
PSS839531
PSS839521
PSS839566
PSS839526
PSS839511
PSS839516
PSS839537
PSS839507
PSS839532
PSS839522
PSS839567
PSS839527
PSS839512
PSS839517
PSS839502
PSS839538
PSS839508
PSS839613
PSS839533
PSS839523
PSS839568
PSS839528
PSS839513
PSS839518
PSS839503
PSS839540
PSS839510
PSS839615
PSS839535
PSS839525
PSS839570
PSS839530
PSS839515
PSS839520
PSS839505
PSS845304
PSS845308
PSS845305
PSS845309
PSS832711
PSS837511
PSS832712
PSS837512
PSS832713
PSS837513
PSS832715
PSS837515
PSS839571
PSS839651
PSS839572
PSS839652
PSS839573
PSS839653
PSS839575
PSS839655
PSS837812
PSS837813
PSS837815
PSS839643
PSS839645
Ion Exchange
S5 SAX
S5 SCX
Mixed Mode
S5 OD/CN
Cartridge Columns
4.6 mm
x 50 mm
4.6 mm
x 60 mm
4.6 mm
x 100 mm
4.6 mm
x 150 mm
4.6 mm
x 30 mm
4.6 mm
x 50 mm
4.6 mm
x 100 mm
4.6 mm
x 150 mm
PSS832111
PSS833411
PSS832211
PSS833111
PSS832911
PSS832311
PSS833711
PSS832411
PSS832011
PSS839853
PSS839852
PSS832112
PSS833412
PSS832212
PSS833112
PSS832912
PSS832312
PSS833712
PSS832412
PSS832012
PSS832113
PSS833413
PSS832213
PSS833113
PSS832913
PSS832313
PSS833713
PSS832413
PSS832013
PSS830065
PSS830068
PSS839546
PSS839551
PSS839581
PSS839576
PSS839556
PSS839591
PSS839561
PSS839541
PSS839547
PSS839587
PSS839552
PSS839582
PSS839577
PSS839557
PSS839592
PSS839562
PSS839542
PSS839548
PSS839588
PSS839553
PSS839583
PSS839578
PSS839558
PSS839593
PSS839563
PSS839543
3.0 mm
x 125 mm
3.0 mm
x 250 mm
4.0 mm
x 125 mm
4.0 mm
x 250 mm
PSS838529
PSS838530
PSS838533
PSS838535
PSS838531
PSS838534
PSS838522
PSS838523
PSS838525
PSS838526
PSS838524
PSS838521
PSS845553
PSS845551
PSS845555
PSS845557
PSS845559
PSS845565
PSS845561
PSS845563
PSS845549
PSS845554
PSS845552
PSS845556
PSS845558
PSS845560
PSS845566
PSS845562
PSS845564
PSS845550
PSS845567
PSS845569
PSS845568
PSS845570
Ion Exchange
S5 SAX
S5 SCX
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5 m
Chemistry
S5 ODS2
S5 ODS1
S5 C8
S5 C6
S5 C1
S5 NH2
S5P (Phenyl)
S5 CN
S5 W (Silica)
10 mm
x 250 mm
20 mm
x 250 mm
PSS831985
PSS830685
PSS831885
PSS831085
PSS832685
PSS831185
PSS830885
PSS830985
PSS830185
PSS831995
PSS830695
PSS831895
PSS831095
PSS832695
PSS831195
PSS830895
PSS830995
PSS830195
PSS832785
PSS837585
PSS832795
PSS837595
10 m
Chemistry
10 mm
x 250 mm
20 mm
x 250 mm
S10 ODS2
S10 ODS1
S10 C8
S10 C6
S10 C1
S10 NH2
S10 P (Phenyl)
S10 CN
S10 W (Silica)
PSS832585
PSS830785
PSS832885
PSS833285
PSS833085
PSS833685
PSS833885
PSS833585
PSS830285
PSS832595
PSS830795
PSS832895
PSS833295
PSS833095
PSS833695
PSS833895
PSS833595
PSS830295
S10 SAX
S10 SCX
PSS833985
PSS837685
PSS833995
PSS837695
Ion Exchange
S5 SAX
S5 SCX
Analytical Chromatography
153
aNaLYTICaL ChROMaTOGRaPhY
Waters Spherisorb Microbore Columns
3 m
Chemistry
1 mm
x 100 mm
1 mm
x 150 mm
S3 ODS2
S3 ODS1
S3 C8
PSS832132
PSS832232
PSS832133
PSS833433
PSS832233
S3 NH2
S3 CN
S3 W (Silica)
PSS832432
PSS832032
PSS832333
PSS832433
PSS832033
5 m
Chemistry
S5 ODS2
S5 ODS1
S5 C8
S5 C6
S5 NH2
S5 CN
S5 W (Silica)
1 mm
x 100 mm
1 mm
x 150 mm
1 mm
x 250 mm
PSS831932
PSS830632
PSS831832
PSS831032
PSS831132
PSS830932
PSS831933
PSS831935
PSS830635
PSS831135
PSS830135
2 mm
x 100 mm
2 mm
x 150 mm
2 mm
x 250 mm
PSS831922
PSS830622
PSS831822
PSS831122
PSS830922
PSS830122
PSS831923
PSS830623
PSS831823
PSS831123
PSS830923
PSS830123
PSS831925
PSS830625
PSS831825
PSS831125
PSS830925
PSS830125
2 mm
x 100 mm
2 mm
x 150 mm
PSS832122
PSS833422
PSS832222
PSS832322
PSS832422
PSS832022
PSS832123
PSS833423
PSS832223
PSS832323
PSS832423
PSS832023
5 m
Chemistry
S5 ODS2
S5 ODS1
S5 C8
S5 NH2
S5 CN
S5 W (Silica)
Qty.
Part No.
2/pk
5/pk
5/pk
2/pk
2/pk
1/pk
1/pk
PSS614100
PSS614103
PSS614104
PSS614102
PSS614101
PSS614108
WAT015674
10/pk
PSS614113
PSS830008
Chemistry
10 x 4.6 mm
3/pk
30 x 4.6 mm
Guard 3/pk
S5 ODS2
S5 ODS1
S5 C8
S5 C6
S5 C1
S5 CN
S5 Phenyl
S5 NH2
S5 W Silica
S5 SAX
S5 SCX
S5 ODSB
PSS830053
PSS830073
PSS830074
PSS830075
PSS830076
PSS830077
PSS830078
PSS830079
PSS830051
PSS830055
PSS830057
PSS830059
PSS839458
PSS839472
PSS839473
PSS839474
PSS839475
PSS839476
PSS839477
PSS839478
PSS839451
PSS839465
PSS839471
PSS839470
154
Analytical Chromatography
Extended Endfitting
for use with 10 mm Guard
PSS 614108
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aNaLYTICaL ChROMaTOGRaPhY
Nova-Pak Packings
T he 4 m small particle size of Nova-Pak bonded phases offers high
resolution as well as more efficient and faster c hromatography. T his
is accomplished by using shorter columns, thereby reducing solvent
consumption or by using longer columns to resolve even the most
complex mixture. Analytical columns with 4 m particle size packings
are available in 75, 150, and 300 mm length steel columns. Steel
cartridge columns with reusable endfittings are available in 50, 100,
150, and 250 mm lengths. Semi-preparative columns with 6 micron
particle size packings give you an unparalleled range of separation
possibilities. Stringent QC procedures in our cGMP manufacturing facility
ensure batc h-to-batc h reproducibility.
Rapid Analysis of Soft Drink Additives and Artificial
Sweeteners on Nova-Pak C18 Column
Column:
Part Number:
Mobile Phase:
Flow Rate:
Detection:
Sample:
Physical Characteristics
Packing
Chemistry
Particle
Size
Particle
Shape
Pore
Size
Carbon
Load
Endcapped
Nova-Pak
C18
C8
Phenyl
CN HP
Silica
4 m
4 m
4 m
4 m
4 m
Spherical
Spherical
Spherical
Spherical
Spherical
60
60
60
60
60
7%
4%
5%
2%
N/A
Yes
Yes
Yes
Yes
N/A
Prep Nova-Pak HR
C18
Silica
6 m
6 m
Spherical
Spherical
60
60
7%
N/A
Yes
N/A
Anticonvulsants
Column:
Part Number:
Sample:
In elution order:
6 Min.
3 Min.
Explosives
Blank
Min.
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Analytical Chromatography
155
aNaLYTICaL ChROMaTOGRaPhY
Nova-Pak Analytical Columns
Particle Size
Dimension
Part No.
4 m
2.1 x 150 mm
3.9 x 75 mm
3.9 x 150 mm
3.9 x 300 mm
4.6 x 150 mm
2.1 x 150 mm
3.9 x 75 mm
3.9 x 150 mm
2.1 x 150 mm
3.9 x 75 mm
3.9 x 150 mm
3.9 x 75 mm
3.9 x 150 mm
3.9 x 300 mm
2.1 x 150 mm
3.9 x 75 mm
3.9 x 150 mm
WAT023655
WAT011670
WAT086344
WAT011695
WAT044375
WAT052735
WAT035877
WAT035876
WAT052740
WAT011675
WAT010656
WAT010270
WAT044245
WAT056920
WAT052745
WAT011680
WAT010025
Nova-Pak C8
4 m
Nova-Pak Phenyl
4 m
Nova-Pak CN HP
4 m
Nova-Pak Silica
4 m
Particle Size
Qty.
Dimension
Part No.
Nova-Pak C18
4 m
Nova-Pak C8
4 m
Nova-Pak Phenyl
4 m
Nova-Pak CN HP
4 m
Nova-Pak Silica
4 m
1/box
3/box
1/box
1/box
3/box
1/box
1/box
1/box
1/box
1/box
3/box
1/box
1/box
1/box
1/box
1/box
3/box
1/box
1/box
1/box
3/box
1/box
1/box
1/box
3/box
3.9 x 50 mm
3.9 x 50 mm
3.9 x 100 mm
3.9 x 150 mm
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
3.9 x 50 mm
3.9 x 100 mm
3.9 x 150 mm
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
3.9 x 50 mm
3.9 x 100 mm
3.9 x 150 mm
3.9 x 150 mm
3.9 x 50 mm
3.9 x 100 mm
3.9 x 150 mm
3.9 x 150 mm
4.6 x 150 mm
4.6 x 250 mm
3.9 x 150 mm
3.9 x 150 mm
WAT052780
WAT052834
WAT052810
WAT036975
WAT037520
WAT052845
WAT052840
WAT052775
WAT052805
WAT036985
WAT054870
WAT052855
WAT052850
WAT052790
WAT052800
WAT036970
WAT054890
WAT052785
WAT052795
WAT044243
WAT044445
WAT044455
WAT044460
WAT036980
WAT054880
WAT037525
156
Analytical Chromatography
Particle Size
Dimension
Part No.
4 m
3.9 x 150 mm
WAT052770
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aNaLYTICaL ChROMaTOGRaPhY
Analytical Kits
Description
Waters Reversed-Phase Scouting Kit
(includes 1 each Nova-Pak C18, C8, and
Phenyl cartridge column)
Waters Method Development Scouting Kit
(includes 1 each Nova-Pak C8, Phenyl, and
CN HP cartridge column)
Waters Method Validation Kit
(includes 3 Nova-Pak C18 columns representing
2 different bonded-silica bulk packing batches)
Dimension
Part No.
3.9 x 50 mm
WAT044360
3.9 x 50 mm
WAT044255
3.9 x 150 mm
WAT052770
Description
Part No.
WAT046905
WAT046910
Description
Particle Size
Dimension
Part No.
4 m
4 m
4 m
4 m
4 m
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
WAT044380
WAT046830
WAT046835
WAT046845
WAT046840
Particle Size
Pore Size
Part No.
4 m
4 m
4 m
4 m
4 m
60
60
60
60
60
WAT015220
WAT035880
WAT020795
WAT020800
WAT020790
Nova-Pak C18
Nova-Pak C8
Nova-Pak Phenyl
Nova-Pak CN HP
Nova-Pak Silica
Guard-Pak Holder
Guard-Pak Holder
WAT088141
www.waters.com
Particle Size
Pore Size
Dimension
Part No.
6 m
60
3.9 x 300 mm
7.8 x 300 mm
19 x 300 mm
WAT038500
WAT025820
WAT025822
Prep Nova-Pak
HR Silica
6 m
60
3.9 x 300 mm
WAT038501
7.8 x 300 mm
19 x 300 mm
WAT025821
WAT025823
Analytical Chromatography
157
aNaLYTICaL ChROMaTOGRaPhY
Bondapak/Bondapak Packings
If your method calls for Bondapak C 18 packings, there is only one
Bondapak C18 packing material. Almost anything can be made once, but
making a packing material reproducible year-to-year for over 30 years
demonstrates true manufacturing and QC capability. Many companies
claim to have columns with Bondapak-like selectivity but not one has
ever passed Waters stringent QC batch test. Waters has set the standard
for reproducible, high-performance c hromatographic columns. Waters
Bondapak columns are still the most widely referenced and requested
columns in the world.
Pore Size
Dimension
Part No.
Bondapak C18
Description
10 m
125
Bondapak C18
15-20 m
125
10 m
10 m
10 m
125
125
125
7.8 x 300 mm
19 x 150 mm
19 x 300 mm
7.8 x 300 mm
19 x 300 mm
7.8 x 300 mm
7.8 x 300 mm
7.8 x 300 mm
WAT084176
WAT088500
WAT025828
WAT025832
WAT025834
WAT084177
WAT084178
WAT084179
Bondapak CN
Bondapak NH2
Bondapak Phenyl
Physical Characteristics
Packing
Bondapak
Bondapak
Bondapak
Bondapak
Bondapak
Chemistry
Particle
Size
Particle
Shape
Pore
Size
Carbon
Load
Endcapped
C18
Phenyl
CN
NH2
C18
10 m
10 m
10 m
10 m
15-20 m
Irregular
Irregular
Irregular
Irregular
Irregular
125
125
125
125
125
10%
8%
6%
3.5%
10%
Yes
Yes
Yes
No
Yes
Bondapak C18
Bondapak C18
Bondapak CN
Bondapak NH2
Bondapak Phenyl
Particle Size
10 m
15-20 m
10 m
10 m
10 m
Pore Size
Dimension
Part No.
125
2.1 x 300 mm
3.9 x 150 mm
3.9 x 300 mm
4.6 x 150 mm
4.6 x 300 mm
3.9 x 300 mm
3.9 x 150 mm
3.9 x 300 mm
3.9 x 150 mm
3.9 x 300 mm
WAT086609
WAT086684
WAT027324
WAT044370
186000925
WAT025875
WAT086688
WAT084040
WAT086680
WAT027198
125
125
125
125
Pore Size
Dimension
Bondapak C18
10 m
125
4.6 x 250 mm
Endfittings
(includes 1 pair of reusable endfittings, 2 c-clips and coupling)
158
10 m
10 m
10 m
10 m
WAT044480
WAT046855
WAT046865
WAT046850
Description
Analytical Chromatography
Part No.
WAT046905
WAT046910
Part No.
Bondapak C18
Bondapak NH2
Bondapak Phenyl
Bondapak CN
WAT052860
Guard-Pak Holder
WAT037525
Guard-Pak Holder
(Requires endfittings)
Particle Size
Part No.
Description
Description
Particle Size
Particle Size
10 m
10 m
10 m
10 m
Pore Size
125
125
125
125
Part No.
WAT088070
WAT026760
WAT026745
WAT026750
WAT088141
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
Delta-Pak Columns
Delta-Pak columns, ideal for the separation of peptides, proteins and
natural products, are based on a highly stable, bonded, endcapped 5 m
or 15 m spherical silica. Delta-Pak columns are available in two different
pore size materials (100 and 300) with a C18 or C4 bonded phase.
Delta-Pak columns are available in different cartridge and column configurations, providing consistent and predictable scale-up capabilities for
milligram-to-gram scale purifications. Also, by having the same chemistry
available on a high-efficiency 5 m particle, you can easily check the purity
of your fractions.
Physical Characteristics
Packing
Delta-Pak
Particle Size
Pore Size
Dimension
Part No.
Delta-Pak C18
5 m
Delta-Pak C4
5 m
300
100
300
100
300
2.1 x 150 mm
3.9 x 150 mm
3.9 x 150 mm
3.9 x 150 mm
3.9 x 150 mm
WAT023650
WAT011795
WAT011793
WAT011796
WAT011794
Particle Size
Pore Size
Dimension
Part No.
Delta-Pak C18
15 m
Delta-Pak C4
15 m
100
300
100
300
100
300
100
300
100
100
300
100
300
100
300
100
300
3.9 x 300 mm
3.9 x 300 mm
7.8 x 300 mm
7.8 x 300 mm
19 x 300 mm
19 x 300 mm
30 x 300 mm
30 x 300 mm
50 x 300 mm
3.9 x 300 mm
3.9 x 300 mm
7.8 x 300 mm
7.8 x 300 mm
19 x 300 mm
19 x 300 mm
30 x 300 mm
30 x 300 mm
WAT011797
WAT011802
WAT011798
WAT011803
WAT011799
WAT011804
WAT011800
WAT011805
WAT011801
WAT011807
WAT011812
WAT011808
WAT011813
WAT011809
WAT011814
WAT011810
WAT011815
Chemistry
Particle
Size
Particle
Shape
Pore
Size
Carbon
Load
Endcapped
C18
C18
5 m
5 m
Spherical
Spherical
100
300
17%
7%
Yes
Yes
C4
C4
5 m
5 m
Spherical
Spherical
100
300
7%
3%
Yes
Yes
C18
C18
15 m
15 m
Spherical
Spherical
100
300
17%
7%
Yes
Yes
C4
C4
15 m
15 m
Spherical
Spherical
100
300
7%
3%
Yes
Yes
www.waters.com
Pore Size
Dimension
Analytical Chromatography
Part No.
WAT035571
WAT022999
159
ANALYTICAL CHROMATOGRAPHY
Delta-Pak Sentry Guard Columns (3.9 x 20 mm) 2/pk
Description
Particle Size
Pore Size
Part No.
5 m
5 m
5 m
5 m
100
300
100
300
WAT046880
WAT046890
WAT046875
WAT046885
Delta-Pak C18
Delta-Pak C18
Delta-Pak C4
Delta-Pak C4
Description
Particle Size
Pore Size
Quantity
Part No.
5 m
5 m
5 m
100
300
300
10/pk
10/pk
10/pk
WAT036870
WAT036875
WAT036865
Delta-Pak C18
Delta-Pak C18
Delta-Pak C4
Guard-Pak Holder
WAT088141
Part No.
WAT046905
WAT046910
Physical Characteristics
Particle Size
Part No.
10 m
WAT046860
Porasil
Packing
Chemistry
Particle
Size
Particle
Shape
Pore
Size
Carbon
Load
Endcapped
Porasil
Porasil
Silica
Silica
10 m
15-20 m
Irregular
Irregular
125
125
N/A
N/A
N/A
N/A
Pore Size
Dimension
Part No.
10 m
125
Porasil
15-20 m
125
3.9 x 150 mm
3.9 x 300 mm
7.8 x 300 mm
19 x 150 mm
19 x 300 mm
3.9 x 300 mm
19 x 300 mm
WAT086692
WAT027477
WAT084175
WAT091648
WAT025829
WAT025874
WAT025835
160
Analytical Chromatography
Porasil
Particle Size
Part No.
WAT046905
WAT046910
www.waters.com
ANALYTICAL CHROMATOGRAPHY
Resolve Columns
T he non-endcapped Resolve packings are significantly different from
Nova-Pak and Bondapak packing materials. T he change in chromatographic
be havior is most commonly noticed wit h polar compounds w hic h are
typically more retained. Basic compounds can be chromatographed using
mobile phase modifiers such as ion-pairing reagents which reduce excessive
tailing. Resolve C 18 and Silica columns are available for applications
requiring high resolution.
Particle Size
Pore Size
Part No.
Resolve C18
Resolve C8
5 m
5 m
90
90
WAT046915
WAT046920
Physical Characteristics
(50 mm and 75 mm long cartridge columns must use the Universal Guard Holder)
Packing
Chemistry
Particle
Size
Particle
Shape
Pore
Size
Carbon
Load
Endcapped
Resolve
Silica
C18
C18
C8
CN
Silica
5 m
5 m
10 m
10 m
10 m
10 m
Spherical
Spherical
Spherical
Spherical
Spherical
Spherical
90
90
90
90
90
90
N/A
10%
10%
5%
3%
N/A
N/A
No
No
No
No
N/A
Description
Part No.
WAT046905
WAT046910
Resolve Columns
Description
Description
Particle Size
Pore Size
Dimension
Part No.
Resolve C18
5 m
90
Resolve Silica
5 m
90
3.9 x 300 mm
3.9 x 150 mm
3.9 x 150 mm
WAT011740
WAT085711
WAT086016
Resolve C18
Resolve Silica
Resolve C8
Resolve CN
Particle Size
Pore Size
Part No.
10 m
10 m
10 m
10 m
90
90
90
90
WAT085824
WAT085825
WAT026755
WAT085826
Guard-Pak Holder
WAT088141
www.waters.com
Description
Part No.
Analytical Chromatography
715001531
715002531
161
aNaLYTICaL ChROMaTOGRaPhY
RSpak D Series
Column Type
DS-613
DE-613
DC-613
Base Polymer
Functional Group
Polystyrene
Polymethacrylate
Polystyrene
None
None
Sulfonated
Flow Rate:
Detection:
Sample:
30 Min.
Sample:
Solvent:
Local anesthetics
44% acetonitrile, 56% water,
1% ammonium hydroxide (pH 11)
3 mL/min
220 nm
Flow Rate:
Detection:
2
3
1
2
1
2
B: DS - 613
E: C18
18 min.
Above is a comparison of the separation of three local anesthetics using RSpak columns DE-613, DS-613,
D18-613 and D4-613. Mobile phase contains ammonium hydroxide at pH 11. The compounds separated are
lidocaine, procaine and tetracaine. The pK values of the anesthetics are greater than 9.0, a mobile phase with a
high pH is therefore needed to deprotonate these compounds. Once deprotonated, they are easily separated by a
reversed-phase mechanism. Eluents are simple and easier to reproduce. Compare these to Figure E, a separation
performed on a silica-based C18 column using an ion pairing reagent.
162
Analytical Chromatography
Dimension
Part No.
6 x 150 mm
6 x 150 mm
6 x 150 mm
4.6 x 10 mm
4.6 x 10 mm
4.6 x 10 mm
WAT034220
WAT034221
WAT034223
WAT034224
WAT034225
WAT034227
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aNaLYTICaL ChROMaTOGRaPhY
Guard Products
Guide to Guard Column Selection and Use
Guard columns are used primarily to protect expensive analytical and
preparative columns by removing particulate and strongly retained
sample components whic h ultimately decrease the lifetime of the
analytical column. Although economy alone is a persuasive argument
for the use of guard columns, the need for a long, stable life in the
analytical column in order to obtain reliable and reproducible results
is per haps even more important.
Guard columns are installed between the injector and the analytical
column of a liquid chromatographic system. Guard columns may also be
used to perform automated, on-line sample cleanup. In situations where
a guard column is not needed, or where no suitable guard column is
available, an in-line filter should be used to protect the analytical column
from microparticulate contamination. T he in-line filter is placed between
the injector and the column.
Although larger guard columns would have the capacity to trap impurities from more sample injections, excessive band broadening would result.
Waters offers two configurations for guard columnsthe Sentry guard
holders and Guard-Pak holder. T he Sentry guard holder is available in two
designs: one to be used as an integrated part of the cartridge column with
reusable endfittings; the other for use with any HPLC column. Both are
installed without tools. T he Guard-Pak holder is a convenient and inexpensive device since you can use prepacked Guard-Pak inserts matched to the
chemistry of your analytical column.
Positioning of a Guard Column in an LC System
Solvent
Delivery
System
Detector
Injector
Guard Column
or Inline Filter
HPLC Column
Flow
www.waters.com
Part No.
Analytical Chromatography
WAT046905
WAT046910
WAT097958
WAT097954
WAT022681
186000262
163
ANALYTICAL CHROMATOGRAPHY
Sentry Guard Columns
Dimension
2.5 m
3.5 m
5 m
XBridge C18
2.1 x 10 mm
3.0 x 20 mm
4.6 x 20 mm
2.1 x 10 mm
3.0 x 20 mm
4.6 x 20 mm
2.1 x 10 mm
3.0 x 20 mm
4.6 x 20 mm
2.1 x 10 mm
3.0 x 20 mm
4.6 x 20 mm
186003056
186003057
186003058
186003074
186003075
186003076
186003065
186003066
186003067
186003359
186003360
186003361
186003059
186003060
186003061
186003077
186003078
186003079
186003068
186003069
186003070
186003362
186003363
186003364
186003062
186003063
186003064
186003080
186003081
186003082
186003071
186003072
186003073
186003366
186003367
186003368
XBridge C8
XBridge Shield RP18
XBridge Phenyl
Description
Dimension
2.5 m
3.5 m
5 m
SunFire C18
2.1 x 10 mm
3.0 x 20 mm
4.6 x 20 mm
2.1 x 10 mm
3.0 x 20 mm
4.6 x 20 mm
186003395
186003405
186003413
186003396
186003406
186003414
186002530
186002681
186002682
186002708
186002718
186002727
186002536
186002683
186002684
186002713
186002722
186002733
Dimension
3 m
5 m
Atlantis T3
2.1 x 10 mm
3.9 x 20 mm
4.6 x 20 mm
2.1 x 10 mm
2.1 x 20 mm
3.9 x 20 mm
4.6 x 20 mm
2.1 x 10 mm
3.9 x 20 mm
4.6 x 20 mm
186003756
186003757
186003758
186001377
186001381
186001313
186001321
186002005
186002021
186002023
186003759
186003760
186003761
186001379
186001383
186001315
186001323
186002006
186002022
186002024
Atlantis dC18
Atlantis HILIC
164
Dimension
3.5 m
5 m
2.1 x 10 mm
2.1 x 20 mm
3.0 x 20 mm
3.9 x 20 mm
2.1 x 10 mm
2.1 x 20 mm
3.0 x 20 mm
3.9 x 20 mm
2.1 x 10 mm
2.1 x 20 mm
3.0 x 20 mm
3.9 x 20 mm
2.1 x 10 mm
2.1 x 20 mm
3.0 x 20 mm
3.9 x 20 mm
186000632
186000636
186000640
186000644
186000633
186000637
186000641
186000645
186000634
186000638
186000642
186000646
186000635
186000639
186000643
186000647
186000648
186000652
186000656
186000660
186000649
186000653
186000657
186000661
186000650
186000654
186000658
186000662
186000651
186000655
186000659
186000663
Analytical Chromatography
Particle Size
Part No.
2.1 x 10 mm
3.9 x 20 mm
2.1 x 10 mm
3.9 x 20 mm
2.1 x 10 mm
3.9 x 20 mm
2.1 x 10 mm
3.9 x 20 mm
2.1 x 10 mm
3.9 x 20 mm
2.1 x 10 mm
3.9 x 20 mm
3.5 m
5 m
3.5 m
5 m
3.5 m
5 m
3.5 m
5 m
3.5 m
5 m
3.5 m
5 m
WAT106127
WAT054225
WAT106128
WAT054250
186000169
186000107
WAT106129
WAT200675
186000275
186000284
186000198
WAT106166
Dimension
Particle Size
Part No.
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
4 m
4 m
4 m
4 m
4 m
WAT044380
WAT046830
WAT046835
WAT046840
WAT046845
Dimension
Particle Size
Part No.
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
10 m
10 m
10 m
10 m
WAT044480
WAT046855
WAT046865
WAT046850
Dimension
Particle Size
Part No.
3.9 x 20 mm
10 m
WAT046860
SunFire C8
Symmetry C18
Symmetry C18
Symmetry C8
Symmetry C8
SymmetryShield RP18
SymmetryShield RP18
SymmetryShield RP8
SymmetryShield RP8
Symmetry300 C4, 300
Symmetry300 C4, 300
Symmetry300 C18, 300
Symmetry300 C18, 300
Dimension
Nova-Pak C18
Nova-Pak C8
Nova-Pak Phenyl
Nova-Pak CN HP
Nova-Pak Silica
Dimension
Particle Size
Part No.
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
3.9 x 20 mm
5 m
5 m
5 m
5 m
WAT046880
WAT046890
WAT046875
WAT046885
Dimension
Particle Size
Part No.
3.9 x 20 mm
4 m
WAT046895
Dimension
Particle Size
Part No.
Resolve C18
Resolve C8
3.9 x 20 mm
3.9 x 20 mm
5 m
5 m
WAT046915
WAT046920
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aNaLYTICaL ChROMaTOGRaPhY
Guard-Pak: Inexpensive Column Protection
Guard-Pak Holder
Guard-Pak Inserts
Description
Part No.
Guard-Pak Holder
Guard-Pak Holder Connector
WAT088141
WAT080046
Description
Nova-Pak C18
Nova-Pak C8
Nova-Pak Phenyl
Nova-Pak Silica
Nova-Pak CN HP
Bondapak C18
Bondapak NH2
Bondapak Phenyl
Bondapak CN
Resolve C18
Resolve Silica
Resolve C8
Resolve CN
Delta-Pak C18
Delta-Pak C18
Delta-Pak C4
Guard-Pak In-Line Filters
Qty.
Particle Size
Pore Size
Part No.
10/pk
10/pk
10/pk
10/pk
10/pk
10/pk
10/pk
10/pk
10/pk
10/pk
10/pk
10/pk
10/pk
10/pk
10/pk
10/pk
5/pk
4 m
4 m
4 m
4 m
4 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
5 m
5 m
5 m
60
60
60
60
60
125
125
125
125
90
90
90
90
100
300
300
WAT015220
WAT035880
WAT020795
WAT020790
WAT020800
WAT088070
WAT026760
WAT026745
WAT026750
WAT085824
WAT085825
WAT026755
WAT085826
WAT036870
WAT036875
WAT036865
WAT032472
Universal Sentry
Guard Holder
Cartridge Columns
Cartridge End-Fittings
2.1 x 50 mm, 2.1 x 100 mm, 2.1 x 150 mm, 2.1 x 250 mm use Part No. 700000117
3.0 x 50 mm, 3.0 x 100 mm, 3.0 x 150 mm, 3.0 x 250 mm use Part No. WAT037525
3.9 x 50 mm, 3.9 x 100 mm, 3.9 x 150 mm, 3.9 x 250 mm use Part No. WAT037525
4.6 x 50 mm, 4.6 x 100 mm, 4.6 x 150 mm, 4.6 x 250 mm use Part No. WAT037525
Integrated Sentry Guard Holder to go with
cartridge end-fittings (see above)
Sentry Guard
Column
www.waters.com
Analytical Chromatography
165
aNaLYTICaL ChROMaTOGRaPhY
Use Low UV PIC to analyze ionic compounds at low UV (200 nm). Low UV
PIC is specially formulated to eliminate background interference in the low
UV range and provides:
Alkaloids
Cephaeline
Emetine
Ergot alkaloids
Analgesics/Antipyretics
APC
Aspirin
Codeine
Indomethacin
Meperidine
Methadone
Morphine
Propoxyphene
Antibiotics
Cephalosporins
Penicillins
Antihistamines
Chlorpheniramine
Diphenhydramine
Methapyrilene
Phenimdamine
Pheniramine
Pyrilamine
Tripelennamine
PIC Reagent
B
B
B
A
A
B
A
B
B
B
B
A or B
A or B
B
B
B
B
B
B
B
Compound
or Class
Antitussives
B Vitamins
Calcium Leucovorin
FD&C Dyes
Folic Acid
Germicides
Benzalkonium Chloride
Cetylpyridinium
Imides
Methotrexate
Petroleum Sulfonates
Surfactants
Quaternary amines
Sulfonates
Sympathomimetics
(Decongestants)
Amphetamine
Ephedrine
Epinephrine
Isoproterenol
Naphazoline
Nylidrin
Phenylephrine
Phenylpropanolamine
166
Analytical Chromatography
Sample:
Compound
or Class
Use PIC B for rapid separation of bases. Quaternary amines, another class
of compounds that is usually separated by ion-exchange chromatography,
can be more easily separated by Paired-Ion Chromatography employing one
of the PIC Reagent B series.
Use Reagent PIC D-4 as a mobile-phase modifier when chromatographing
basic pharmaceutical compounds to improve peak shape and reduce
retention times.
Use PIC A for rapid separation of acids. FD&C sulfonic acid dyes are classic
examples of compounds that are conventionally separated by ion-exchange
chromatography. Using PIC Reagent A and reversed-phase chromatography,
the sulfonic acid dyes and their impurities can be separated easier, faster,
and more reliably.
Flow Rate:
Detection:
PIC Reagent
B
B5/B7
A
A
A
B
B
A
A
A
B
A
B
B
B
B
B
B
B
B
Workable
UV Range in
Mobile Phase (nm)
PIC A (tetrabutylammonium hydrogen sulfate)
Low UV PIC A
PIC A (tetrabutylammonium phosphate
PIC B5 (pentane sulfonic acid)
Low UV PIC B5
PIC B5
PIC B6 (hexane sulfonic acid)
Low UV PIC B6
PIC B6
PIC B7 (heptane sulfonic acid)
Low UV PIC B7
PIC B7
PIC B8 (octane sulfonic acid)
Low UV PIC B8
PIC B8
PIC Sample Kit (One vial each: PIC A,
B5, B6, B7 and B8)
PIC Reagent D4 (dibutylammonium phosphate)
Quantity
(Vials/
Pkg)
Part No.
200+
240+
5
5
WAT084189
WAT085101
200+
240+
5
5
WAT084198
WAT085110
200+
240+
5
5
WAT084199
WAT085140
200+
240+
5
5
WAT084282
WAT085103
200+
240+
5
5
WAT084283
WAT085142
215+
5
5
WAT085144
WAT085466
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
Reversed-Phase
Reversed-phase is best suited to the separation of mixtures containing
monosaccharides and oligosaccharides up to a degree of polymerization
(DP) of 8 as applies to monitoring acid or enzyme hydrolysates of large
oligosacc harides. T his mode is also amenable for the separation of
derivatized sugars.
Size Exclusion
Gel-Permeation Chromatography (GPC) is used to determine the molecular
weight distribution of food polymers such as starches and gums. T he Waters
Ultra hydrogel columns are well suited for this application. T he Shodex
KS Series (7 m) and robust S Series (10 m) packings are based on a
sulfonated styrene divinylbenzene polymer. Designed to analyze and
c haracterize water-soluble polymers, they are particularly well suited
for mono-, di- and polysacc harides. T he KS-800 Series is also useful
for separation of intermediate and small DP sugars.
Ion Exchange
Partition
Partition chromatography is best suited for low molecular weight sugars
such as mono-, di-, and trisaccharides. T his mode uses a covalently bonded
amino packing in a column such as our Carbohydrate Analysis column, or
a dynamically-coated silica cartridge employing Silica Amine Modification
(SAM) reagents such as SAM I. Typical mobile phases range from 65-85%
acetonitrile in water, and the separations are carried out at temperatures
ranging from ambient to 70 C.
www.waters.com
Analytical Chromatography
167
aNaLYTICaL ChROMaTOGRaPhY
Typical Applications for Sugar and Carbohydrate Columns
Cartridge/Column
Mode
Eluent
Application
Elution Order
Partition
65 to 85% CH3CN:H2O
ambient to 70 C
Partition
70 to 80% CH3CN:H2O
0.1% SAM I ambient
Ion exchange/
size exclusion
H2O
75-95 C
Ion exchange/
size exclusion
0.01N H3P04
50-60 C
Dextro-Pak
Reversed-phase
H2O
ambient
Hydrolysed syrups
derivatized sugars
KS-800 series
Size exclusion
18
Cation
H = H+
C = Ca2+
P = Pb2+
Z = Zn2+
% cross
linkage
Pore Size
1 = 20
2 = 50
3 = 100
4 = 500
5 = 1000
0 - Gel type
1 - Semimacropore gel
2 - Permanent pore gel
Example:
S
Sugar column
168
Ca2+
10
20 pore size
Analytical Chromatography
semimacropore gel
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
Retention Times for Sugar and Carbohydrates
Column Type
Carbohydrate
Analysis
Sugar-Pak
Dextro-Pak
Silica-Pak
KS-801
SC-1011
SP-0810
SH-1011
Column Temp:
Ambient
90 C
Ambient
75 C
80 C
80 C
60 C
Mobile Phase:
80:20
Acetonitrile:
Water
2 mL/min
Water
Water
Water
Water
Water
Water
0.5 mL/min
1 mL/min
Ambient
SAM I Reagent
75:25
Acetonitrile:
Water
4 mL/min
1 mL/min
1 mL/min
1 mL/min
1 mL/min
8.4
9.2
7.7
9.8
9.0
11.1
10.7
10.8
9.3
13.5
11.1
16.5
7.9
8.6
7.4
7.7
7.5
8.0
5.4
7.9
5.5
9.3
10.6
12.7
20.8
9.0
8.9
8.2
9.6
7.6
11.5
11.2
10.4
9.0
8.7
6.4
6.6
7.1
9.3
13.6
15.3
8.1
8.6
9.9
11.4
13.2
6.6
6.8
7.2
8.3
7.5
7.9
11.3
16.5
11.4
8.7
7.3
13.3
7.0
13.5
7.0
9.8
11.5
9.1
6.1
8.3
13.5
13.2
8.2
5.7
6.4
6.4
6.6
13.0
8.1
7.5
10.3
19.9
22.3
10.2
7.1
8.0
8.1
8.5
20.5
9.7
5.1
7.3
7.9
7.6
6.7
4.9
Flow Rate:
Compound
2-Deoxygalactose
2-Deoxyribose
2-Deoxy-glucose
Adonitol (Ribitol)
Arabinose
Arabitol
Butanol-1
Cellobiose
Deuterium Oxide
Digitose
Digitoxose
Dulcitol
Ethylene Glycol
Ethanol
Fructose
Fucose
Galactose
Glucoheptose
Glucose
Glucose-6-phosphate
Glycerol
Inositol
Isomaltitol
Isomaltose
Lactose
Lactulose
Lyxose
Maltitol
Maltoheptaose
Maltohexaose
Maltopentaose
Maltose
Maltotetraose
Maltotriose
Maltotriol
Mannitol
Mannose
Melibiose
Melizitose
meso-Erythritol
Methanol
myo-Inositol
Pannose
Pentaerythritol
Propanol-2
Raffinose
Rhamnose
Ribose
Sorbitol
Sorbose
Stachyose
Sucrose
Trehalose
Turanose
Xylitol
Xylose
Xylulose
www.waters.com
3.9
4.8
4.8
8.8
11.0
10.4
3.4
2.9
3.0
1.5
1.8
1.8
N/R
15.3
18.8
6.7
N/R
3.7
4.1
8.3
5.7
11.1
N/R
N/R
5.7
4.2
7.7
12.6
12.8
10.1
10.4
9.4
3.5
6.8
2.9
4.1
2.8
1.0
2.0
1.6
2.5
9.3
10.0
7.7
8.1
7.6
7.3
N/R
N/R
8.5
4.8
12.1
2.8
2.1
3.2
2.4
7.2
1.2
8.5
18.0
16.3
4.1
7.1
7.6
10.7
3.0/3.2*
3.8
3.0
4.8
4.0
5.9
5.9
6.0
15.7
6.8
3.5
4.1
4.7
5.9
N/R
6.0
4.4/4.7*
7.8
5.3
7.0
6.4
21.6
12.2
9.6
7.0
2.9
2.9
3.1
2.2
2.2
5.3
7.2
7.6
5.9
5.1
N/R
12.5
3.9
5.9
5.0
12.6
6.0
9.5
14.9
14.2
9.4
5.4
6.8
N/R
7.0
3.3
3.1
2.9
2.8
6.3
4.4
N/R
N/R
3.6
6.9
5.6
N/R
11.9
5.1
4.3
3.3
14.3
9.3
10.6
2.9
2.9
3.1
7.5
1.4
2.2
17.5
3.4
1.8
1.6
5.2
7.3
9.1
7.5
7.4
4.8
5.8
5.7
8.0
7.7
5.4
5.3
5.5
5.8
6.1
5.9
11.1
8.3
6.4
6.0
10.1
9.1
8.4
7.6
7.1
7.8
6.9
6.6
6.6
5.6
5.8
7.4
6.1
4.6
4.6
4.7
5.6
4.9
5.1
5.4
7.4
6.9
5.6
5.1
8.4
5.6
5.8
8.2
7.0
N/R
*
Blank
Analytical Chromatography
169
ANALYTICAL CHROMATOGRAPHY
Description
High-Performance Carbohydrate cartridge column
(requires endfittings)
Endfittings (End Connector Kit)
(contains 1 pair endfittings, and c-clips)
Sentry High-Performance Carbohydrate
guard column (2/pk)
Sentry Integrated Guard Holder
(for Waters cartridge columns)
Dimension
Part No.
4.6 x 250 mm
WAT044355
Exclusion Limit
103
KS-801
KS-802
KS-804
KS-800P pre-column
1x
1 x 104
4 x 105
Dimension
Part No.
8 x 300 mm
8 x 300 mm
8 x 300 mm
6 x 50 mm
WAT034276
WAT034277
WAT034279
WAT034282
Exclusion
Limit (PEO)
Dimension
Part No.
120
250
500
1000
2000
5 x 103
8 x 104
4 x 105
1 x 106
7 x 106
7.8 x 300 mm
7.8 x 300 mm
7.8 x 300 mm
7.8 x 300 mm
7.8 x 300 mm
WAT011520
WAT011525
WAT011530
WAT011535
WAT011540
7 x 106
5 x 103
7.8 x 300 mm
7.8 x 300 mm
WAT011545
WAT011550
WAT011565
WAT011570
Column
Ultrahydrogel 120
Ultrahydrogel 250
Ultrahydrogel 500
Ultrahydrogel 1000
Ultrahydrogel 2000
WAT037525
3.9 x 20 mm
WAT046895
WAT046905
Dimension
Part No.
Sugar-Pak 1 column
Sugar-Pak 1 Guard-Pak inserts, 10/pk
6.5 x 300 mm
WAT085188
WAT015209
3.9 x 300 mm
WAT084038
SC-1011 column
SP-0810 column
SC-1011P pre-column
SP-0810P pre-column
8 x 300 mm
8 x 300 mm
6 x 50 mm
6 x 50 mm
WAT034238
WAT036954
WAT034244
WAT034245
Silica-Pak HP cartridge, 4 m
Silica-Pak cartridge, 10 m
8 x 100 mm
8 x 100 mm
WAT010932
WAT010935
WAT010873
170
8 x 100 mm
Analytical Chromatography
WAT085650
www.waters.com
aNaLYTICaL ChROMaTOGRaPhY
Pesticide Analysis
W hen used with the Waters Carbamate Analysis System, the Carbamate
Analysis column provides a guaranteed analysis of the carbamate pesticides
that exceeds the sensitivity required by AOAC Method 985.23.
Carbamate Analysis
Column:
Part Number:
Mobile Phase:
Flow Rate:
Sample:
Injection Volume:
Post Column
Addition:
Detector:
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
Aldicarb sulfoxide
Aldicarb sulfone
Oxamyl
Methomyl
3-Hydroxycarbofuran
Aldicarb
Propoxur
Carbofuran
Carbaryl
1-Naphthol
Methiocarb
5
9
Sample
1.
2.
3.
4.
5.
6.
7.
8.
9.
Capric Acid
Lauric Acid
Myristic Acid
Palmitic Acid
Stearic Acid
Nonadecanoic Acid
Arachidic Acid
Heneicosanoic Acid
Behenic Acid
1
7
2
6
11
10
5 Min.
20 Min.
www.waters.com
3 4
Dimension
Particle Size
Part No.
3.9 x 150 mm
4 m
WAT011690
Column
Carbamate Analysis
Dimension
Part No.
3.9 x 150 mm
WAT035577
Analytical Chromatography
171
aNaLYTICaL ChROMaTOGRaPhY
Part Number:
System:
Eluent A:
Eluent B:
Gradient:
Flow Rate:
Injection:
Sample:
UV @ 254 nm
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
Naphthalene - 20 ppm
Acenaphthylene - 40 ppm
Acenaphthene - 20 ppm
Fluorene - 4 ppm
Phenanthrene - 2 ppm
Anthracene - 2 ppm
Fluoranthene - 4 ppm
Pyrene - 2 ppm
Benzo(a)anthracene - 2 ppm
Chrysene- 2 ppm
Benzo(b)fluoranthene - 4 ppm
Benzo(k)fluoranthene - 2 ppm
Benzo(a)pyrene - 2 ppm
Dibenzo(a,h)anthracene - 4 ppm
Benzo(g,h,I)perylene - 4 ppm
Indeno(1,2,3-cd)pyrene - 2 ppm
11
10
1
5
4
Waters PAH columns are optimized for the HPLC analysis of PAHs. T he
c hromatogram (top right) shows 16 PAH compounds, listed as target
pollutants by the U.S. EPA. T he Waters PAH columns ac hieve baseline
resolution and excellent peak symmetry for all 16 target analytes in
less than 25 minutes, while employing a simple water:acetonitrile
binary gradient. T he resolving power of the PAH Columns provides
superior peak identification and quantitation for PAHs.
Florida Administrative Code 17.700 includes 2 additional compounds
(1-methyl naphthalene and 2-methyl naphthalene ) in addition to the 16
compound EPA 610 mix that we currently use to show the proficiency of
Waters instrumentation to analyze PAH compounds (bottom right). T he new
Waters PAH columns resolve these two compounds along with the other 16.
Waters PAH columns come in seven different dimensions (including a
capillary format), and two particle sizes. Eac h column comes with a
complete Certificate of Analysis backed by a world-class ISO 9002
registered documentation trail.
7
12
13
10
12
14
14
16
18
20
15
22
16
24 Min.
Part Number:
Eluent A:
Eluent B:
Gradient:
Flow Rate:
Injection:
Sample:
UV @ 254 nm
Reference:
1
Peaks :
12
Peaks :
15
11
4*
9 10
14
Naphthalene - 20 ppm
Acenaphthylene - 40 ppm
1-methyl naphthalene - 25 ppm
2-methyl naphthalene - 25 ppm
Acenaphthene - 20 ppm
Fluorene - 4 ppm
Phenanthrene - 2 ppm
Anthracene - 2 ppm
Fluoranthene - 4 ppm
Pyrene - 2 ppm
Benzo(a)anthracene - 2 ppm
Chrysene - 4 ppm
Benzo(b)fluoranthene - 4 ppm
Benzo(k)fluoranthene - 2 ppm
Benzo(a)pyrene - 2 ppm
Dibenzo(a,h)anthracene - 4 ppm
Benzo(g,h,I)perylene -4 ppm
Indeno(1,2,3-cd)pyrene -2 ppm
13
2
3*
1.
2.
3*.
4*.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
16
11
13
15
17
19
21
17
18
23
25 Min.
172
Dimension
Particle Size
Part No.
4.6 x 250 mm
4.6 x 150 mm
4.6 x 50 mm
3.0 x 250 mm
2.1 x 250 mm
2.1 x 150 mm
0.32 x 150 mm
5 m
5 m
3 m
5 m
5 m
5 m
5 m
186001265
186001264
186001260
186001263
186001262
186001261
186001259
Analytical Chromatography
www.waters.com
ANALYTICAL CHROMATOGRAPHY
Injection Volume:
Detection:
Dimension
Particle Size
Part No.
3.9 x 150 mm
4 m
WAT086344
22 Min.
Effective Molecular
Weight Range
Dimension
Packed in
Toluene
Packed in
THF
50-1,500
100-10,000
7.8 x 300 mm
7.8 x 300 mm
WAT085500
WAT085501
WAT010570
WAT010571
www.waters.com
Sample
Oxalic Acid
Maleic Acid
a- Ketoglutaric Acid
Citric Acid
Tartaric Acid
Pyruvic Acid
trans-Aconitic Acid
Glyoxylic Acid
Malic Acid
Malonic Acid
Citraconic Acid
Succinic Acid
Glycolic Acid
Itaconic
Lactic Acid
Retention Time
5.20
5.80
5.90
6.20
6.55
6.65
6.95
7.00
7.05
7.07
7.20
8.00
8.40
8.50
8.60
Column
Fast Fruit Juice Analysis
Fast Fruit Juice Guard-Pak Inserts 10/pk
IC-Pak Ion Exclusion
SH-1011
SH-1011P pre-column
KC-811
KC-811 pre-column
Sample
Retention Time
- Hydroxypropionic Acid
D-Glucuronic Acid
Fumaric Acid
Formic Acid
Acetic Acid
Adipic Acid
Levulinic Acid
Mesaconic Acid
Pyroglutamic Acid
Propionic Acid
Acrylic Acid
Pivalic Acid
Methacrylic Acid
trans-Crotonic Acid
8.60
8.65
8.95
9.20
9.80
9.80
10.00
10.40
10.70
11.25
11.60
14.05
14.10
15.65
Dimension
Part No.
7.8 x 150 mm
WAT010639
WAT015207
WAT010290
WAT034236
WAT034243
WAT034298
WAT035501
7.8 x 300 mm
8 x 300 mm
6 x 50 mm
8 x 300 mm
6 x 50 mm
Analytical Chromatography
173
ANALYTICAL CHROMATOGRAPHY
Ion Analysis
Waters offers an array of products for ion c hromatography as well as
innovative capillary electrophoresis products for ion analysis (see the
Capillary Electrophoresis section). In ion c hromatography, anions and
cations are typically measured in two separate analyses. T he columns
offered for eac h type of analysis are briefly described below.
1.
2.
3.
4.
5.
6.
7.
8.
Fluoride
Bicarbonate
Chloride
Nitrite
Bromide
Nitrate
Phosphate
Sulfate
1ppm
2ppm
4ppm
4ppm
4ppm
6ppm
5ppm
3
4
1
6
5
8
7
2
15 Min.
5
1
1.
2.
3.
4.
5.
6.
7.
8.
Fluoride
Bicarbonate
Chloride
Nitrite
Bromide
Nitrate
Phosphate
Sulfate
1ppm
2ppm
4ppm
4ppm
4ppm
6ppm
5ppm
7
2
Column:
Part Number:
Sample:
Eluent:
Detection:
1.
2.
3.
4.
5.
6.
7.
8.
Fluoride
Bicarbonate
Chloride
Nitrite
Bromide
Nitrate
Phosphate
Sulfate
1ppm
2ppm
4ppm
4ppm
4ppm
6ppm
5ppm
15 Min.
IC-Pak Anion HR (high resolution) columns are packed with smaller 6 m particles.
This column is designed for the most demanding separations where the columns high efficiency
gives enhanced resolution and outstanding sensitivity.
2
1
6
7
22 Min.
IC-Pak Anion HC (high capacity) columns use the same packing material as the IC-Pak Anion columns
and are best suited for applications where there are orders of magnitude differences in the concentrations
of the analytes. This is often the case with water and soil samples where you may have to measure ppb levels of
phosphate in the presence of ppm levels of chloride and nitrate. The IC-Pak Anion HC column gives
excellent resolution, even with injection volumes up to 500 L.
Description
IC-Pak Anion
IC-Pak Anion HR
IC-Pak Anion HC
IC-Pak Anion Guard-Pak Kit
(Guard-Pak Holder and 5 inserts)
IC-Pak Anion Concentrator Inserts*, 5/pk
IC-Pak Anion Guard-Pak Inserts*, 5/pk
Guard-Pak Holder
Dimension
Part No.
4.6 x 50 mm
4.6 x 75 mm
4.6 x 150 mm
WAT007355
WAT026765
WAT026770
WAT007357
WAT007358
WAT010551
WAT088141
174
Analytical Chromatography
www.waters.com
ANALYTICAL CHROMATOGRAPHY
Ion Analysis
Part Number:
Eluent:
Flow Rate:
Injection:
Detection:
Dimension
Part No.
3.9 x 150 mm
WAT036570
WAT044250
WAT007354
WAT007356
WAT010565
4.6 x 50 mm
4.6 x 50 mm
IC-Pak C M/D
Method:
Column:
Description
1
23
Waters IC-Pak Ion Exclusion columns are used for the analysis of
weak acid anions such as fluoride and short chain organic acids from
formate to butyrate.
4
5
Description
IC-Pak Ion Exclusion column
IC-Pak Ion Exclusion column
IC-Pak Ion Exclusion Guard-Pak inserts*, 10/pk
Guard-Pak Holder
Standards
0.54 ppm
1.0 ppm
2.0 ppm
4.0 ppm
2.0 ppm
3.0 ppm
Dimension
Part No.
7.8 x 150 mm
7.8 x 300 mm
WAT010295
WAT010290
WAT020770
WAT088141
Lanthanide Analysis
Post-column
Reagent:
Part Number:
Sample:
Injection:
Eluent A:
Eluent B:
Column:
Working Standard
1. Cu
=
2. Pb
=
3. Zn
=
4. Ni
=
5. Co
=
6. Cd
=
7. Fe(II) =
8. Mn 8 =
Flow Rate:
Post-column
Reagent:
0.5 ppm
1.0 ppm
0.5 ppm
1.0 ppm
0.5 ppm
1.0 ppm
2.0 ppm
1.0 ppm
Detection:
Delta-Pak C18
Dimension
3.9 x 150 mm
Particle
Size
5 m
10
11
12 13
14
Lu
Yb
Tm
Er
Ho
Dy
Tb
8.
9.
10.
11.
12.
13.
14.
Gd
Eu
Sm
Nd
Pr
Ce
La
4
3
2
1
17 Min.
20 Min.
Description
1.
2.
3.
4.
5.
6.
7.
Working Standard
2 ppm of each
Pore
Size
100
Part No.
WAT011795
Description
Dimension
Resolve C18 *
8 x 100 mm
8 x 10 Cartridge Holder
Particle
Size
Pore
Size
5 m
90
Part No.
WAT084624
WAT082887
www.waters.com
Analytical Chromatography
175
ANALYTICAL CHROMATOGRAPHY
Gas-Chromatography Packings
Waters versatile Porapak gas-c hromatography column packing materials simplify the analysis of a wide variety of complex compounds from
atmospheric gases to organics. Consisting of polymer beads, these unique
packings are chemically and physically stable and have consistent particle
size, porosity and surface area to guarantee analytical reproducibility. T hey
also provide unequaled separation capability with high resolution and low,
constant retention volumes.
Density
(g/cm3)
Single Temp
Prog.
Particle Size
Mesh
Qty.
Part No.
Nonpolar
100-200
0.26
250 C
PS
Nonpolar
100-200
0.26
250 C
Slightly nonpolar
to moderate
500-600
0.34
250 C
QS
Slightly nonpolar
to moderate
500-600
0.34
250 C
Moderate
Polar monomer:
vinyl pyrrolidone
Moderate
Polar monomer:
vinyl pyridine
Polar
Polar monomer:
vinyl pyrrolidone
Highly polar
Polar monomer:
ethyleneglycol
dimethacrylate
450-600
0.32
250 C
300-450
0.35
250 C
250-350
0.41
190 C
225-350
0.39
190 C
50-80
80-100
100-120
50-80
80-100
100-120
50-80
80-100
100-120
50-80
80-100
100-120
50-80
80-100
100-120
50-80
80-100
100-120
50-80
80-100
100-120
50-80
80-100
100-120
20 g
20 g
20 g
20 g
20 g
20 g
26 g
26 g
26 g
26 g
26 g
26 g
24 g
24 g
24 g
26 g
26 g
26 g
29 g
29 g
29 g
31 g
31 g
31 g
WAT027053
WAT027054
WAT027055
WAT027083
WAT027084
WAT027085
WAT027059
WAT027060
WAT027061
WAT027089
WAT027090
WAT027091
WAT027065
WAT027066
WAT027067
WAT027071
WAT027072
WAT027073
WAT027047
WAT027048
WAT027049
WAT027077
WAT027078
WAT027079
Type
Polarity
176
Analytical Chromatography
www.waters.com
PREPaRaTIVE ChROMaTOGRaPhY
Average Bed Density for Conventional Analytical-LC-ColumnSlurry-Packing Procedures vs. Column Aspect Ratio
Packed-Bed Density
[g/cm3]
0.700
Optimum Packed-Bed Density
0.675
4.6 x 100 mm
Analytical LC Columns
0.650
19 x 50 mm
0.625
19 x 100 mm
0.600
50 x 100 mm
Prep LC Columns
0.575
0.550
0
10
15
20
25
OBD Process
Optimum Bed Density Throughout
Packed-bed
Density
Axial-ram-compressed
Slurry Packed
High-pressure
Slurry Packed
Column Inlet
With careful tuning of the packing process for eac h particle type and
column geometry, the Prep OBD design and process results in predictable,
uniform density profiles throughout the column. During the final capping
process, Waters Column Packing Operation follows carefully established
procedures designed not to over-compress or disrupt, in any non-uniform
way, this portion of the bed. Waters scientists have established that too
Column Outlet
muc h axial compression applied at the inlet can break particles, build
bridges, and lower local bed permeability.
T he Prep OBD column is designed to incorporate a pair of speciallydesigned distributors and chemically-inert seals made to prevent leaks at
high operating pressures.
www.waters.com
Preparative Chromatography
177
PREPaRaTIVE ChROMaTOGRaPhY
T he OBD design exhibits exceptional resistance to mechanical chromatographic bed failure and delivers consistent column-to-column performance,
reducing cost through extended lifetimes.
Data from a High-throughput Drug Discovery Laboratory: 7,000 Injections on an XBridge Prep OBD C18 Column, 19 x 50 mm, 5 m
Injection 1
Injection 7,000
100
OBD preparative columns are packed to bed densities which closely match
the equivalent analytical column. T his innovative procedure produces
preparative columns with excellent stability, reproducibility, and efficiency.
SunFire C18
0.1% TFA in Water
0.1% TFA in ACN
Ambient
UV @ 260 nm
Waters AutoPurification System
Compounds
1. Nadolol (100 mg/mL)
2. Metoprolol (100 mg/mL)
3. Propranolol (50 mg/mL)
4.6 x 50 mm
Part Number: 186002557
Total Mass Load: 5 mg
Flow Rate:
Gradient:
1.4 mL/min
Time
(min)
0.0
1.0
6.0
7.0
20 L
Inj. Vol.:
Profile
%A
95
95
30
30
%B
5
5
70
70
-2
100
19 x 50 mm
Part Number: 186002566
Total Mass Load: 85 mg
Flow Rate:
Gradient:
23.9 mL/min
Time
Profile
(min)
%A
0.00
95
1.79
95
6.79
30
7.79
30
340 L
Inj. Vol.:
%B
5
5
70
70
-2
1
178
Preparative Chromatography
6 Min.
www.waters.com
PREPaRaTIVE ChROMaTOGRaPhY
Particle
Size
Pore
Size
Particle
Shape
Carbon
Load
Endcapped
XSelect
C18
Fluoro-Phenyl
Phenyl-Hexyl
5 m
5 m
5 m
130
130
130
Spherical
Spherical
Spherical
15%
10%
14%
Yes
Yes
Yes
XBridge
C18
C8
Shield RP18
Phenyl
HILIC
5, 10 m
5, 10 m
5, 10 m
5 m
5 m
130
130
130
130
130A
Spherical
Spherical
Spherical
Spherical
Spherical
18%
13%
17%
15%
N/A
Yes
Yes
Yes
Yes
No
Packing
SunFire
C18
C8
Silica
5, 10 m
5, 10 m
5, 10 m
100
100
100
Spherical
Spherical
Spherical
16%
12%
N/A
Yes
Yes
Yes
Atlantis
T3
dC18
HILIC
5, 10 m
5, 10 m
5 m
100
100
100
Spherical
Spherical
Spherical
14%
11%
0%
Yes
Yes
No
XTerra MS
C18
C8
5, 10 m
5, 10 m
125
125
Spherical
Spherical
16%
14%
Yes
Yes
XTerra RP
C18
C8
5, 10 m
5, 10 m
125
125
Spherical
Spherical
15%
14%
Symmetry
C18
C8
5, 7 m
5, 7 m
100
100
Spherical
Spherical
SymmetryShield
C18
C8
5, 7 m
5, 7 m
100
100
Symmetry300
C18
C4
5, 7 m
5, 7 m
300
300
Particle
Size
Pore
Size
Particle
Shape
Carbon
Load
Endcapped
6 m
6 m
60
60
Spherical
Spherical
7%
N/A
Yes
N/A
Bondapak
C18
Phenyl
CN
NH2
10 m
10 m
10 m
10 m
125
125
125
125
Irregular
Irregular
Irregular
Irregular
10%
8%
6%
4%
Yes
Yes
Yes
No
Bondapak
C18
15-20 m
37-55 m
125
125
Irregular
Irregular
10%
10%
Yes
Yes
37-55 m
125
Irregular
14%
Yes
Packing
Chemistry
Porasil
Silica
10 m
125
Irregular
N/A
N/A
Yes
Yes
Porasil
Silica
15-20 m
37-55 m
125
125
Irregular
Irregular
N/A
N/A
N/A
N/A
19%
12%
Yes
Yes
Delta-Pak
C18
15 m
100
300
Spherical
Spherical
17%
7%
Yes
Yes
Spherical
Spherical
15%
17%
Yes
Yes
Delta-Pak
C4
15 m
100
300
Spherical
Spherical
7%
3%
Yes
Yes
Spherical
Spherical
9%
3%
Yes
Yes
Prep
C18
Silica
55-105 m
55-105 m
125
125
Irregular
Irregular
11%
N/A
Yes
N/A
No matter how you collect fractions, whether by mass spec, UV, ELS, or
other detector signals, the Waters AutoPurification systems will meet
your needs. T hese systems provide simplicity, scalability, and speed with
a variety of available configurations. T he market leading FractionLynx
Application Manager provides an intuitive interface to manage the
purification process from sample acquisition to reporting.
www.waters.com
Preparative Chromatography
179
PREPaRaTIVE ChROMaTOGRaPhY
Choosing Preparative Columns
Mass Loading
Step 1
Once the analytical separation has been optimized, a loading study on
the analytical column is performed to determine the capacity of the
particular packing material. Because the large-scale separation should
be identical to the small scale separation, the maximum sample load
will be dependent upon the complexity of the analytical separation.
4.6
5
8
13
10
15
25
40
60
Diameter (mm)
19
27
45
90
135
225
30
110
165
225
335
560
50
310
620
930
1550
1.4
20
6.6
100
24
350
60
880
164
2450
Step 2
Determine how much mass you need to purify or isolate.
Step 3
Once the desired purified mass is established, some simple equations
may be used to determine the required column size for purification. In
addition, preparative HPLC system maximum flow rate and back pressure
need to be considered and can limit column size.
T he calculated prep gradient duration is entered into the pumps gradient separation
over the same number of column volumes as was used in the analytical run.
Scale-up factor
Capacity is:
= 17.1
(4.6)2 x 150
(Diameter anal)2
T he calculated flow rate may be used for the larger column to ensure
the same linear velocity of mobile phases as used in the analytical run.
However, reasonable flow rates are based on column diameters. Systems
will be limited by increasing backpressure with increasing column
length and decreasing particle size.
Gradient Duration (GD)
GD (prep) =
Diameter (prep)
Diameter (anal)2
Column/tubing backpressure
Gradient scale-up
Gradient designs
180
Preparative Chromatography
www.waters.com
PREPaRaTIVE ChROMaTOGRaPhY
NEW XSelect Prep OBD Columns
Columns Designed for Isolation and Purification
CSH Tec hnology, especially when combined with the analytical method
development capabilities of UPLC Tec hnology and the industry-leading
column lifetimes offered by Optimum Bed Density (OBD) preparative
columns, may have the greatest benefits to isolation and purification scientists. W hy? CSH Technology offers the highest loading of basic compounds
separated under the volatile, acidic, low pH mobile-phase conditions often
preferred by isolation and purification scientists. T his increased loadability
allows for the use of a narrower (lower volume) preparative column thereby
reducing solvent consumption and fraction volumes.
T he scalability built into ACQUITY UPLC CSH and XSelect columns permits
the use of UPLC Technology as a powerful method development tool for the
isolation and purification scientist. How can c hromatographic separation
platforms that appear to be on opposite ends of the analysis scale spectrum
(microbore vs. preparative) complement eac h other? Again, the answer
lies in scalability and CSH Technologys resistance to pH switching column
performance degradation. Fast, robust, analytical-scale separations
can be easily and quickly developed using UPLC Tec hnology, ACQUIT Y
UPLC CSH columns and simple mobile-phase additives and then straightforwardly scaled and transferred to an XSelect OBD preparative column.
In addition, the purity check of the isolated fraction(s) and/or structural
analysis of isolated impurities can be determined using UPLC/MS.
No other manufacturer can offer the isolation and purification scientist
t he selectivity, loadability and scalability of CSH Tec hnology wit h
the throughput, sensitivity and transferable analytical-scale method
development of UPLC Tec hnology.
Literature
References
ACQUITY UPLC CSH and XSelect HPLC
Columns Brochure, Literature Reference
720003495EN
Charged Surface Hybrid (CSH) Technology and
Its Use in Liquid Chromatography W hite Paper,
Literature Reference 720003469EN
Practical Applications of Charged Surface
Hybrid (CSH) Technology Application Note,
Literature Reference 720003720EN
BEH Technology W hite Paper, Literature
Reference 720001159EN
www.waters.com
Preparative Chromatography
181
PREPARATIVE CHROMATOGRAPHY
LC Conditions
Columns:
Part Number:
Mobile Phase A:
Mobile Phase B:
Gradient:
Compounds
1. Diphenhydramine
2. Oxybutynin
3. Terfenadine
Target Analyte
1
AU
Crude mix
Diphenhydramine = 56.3% purity
1.50
Target Analyte
1
3
0.2
0.4
0.6
0.8
1.0
AU
1.00
1.2
1.4
1.6
1.8
2.0 min
Collected Fraction
0.00
0.0
0.2
AU
0.00
4
0.6
0.8
1.00
1.50
0.00
3
0.50
10 min
Crude mix
Diphenhydramine = 56.3% purity
0.00
0.0
0.2
0.4
0.6
0.8
1.0
AU
AU
1.00
0.50
1.2
1.4
1.6
1.8
2.0 min
0.00
0.0
0.2
0.4
1
2.00
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0 min
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0 min
Collected Fraction
Using CSH Technology throughout the entire process, methods can be developed quickly with ACQUITY UPLC CSH columns and UPLC Technology and then transferred to preparative-scale
OBD XSelect columns for isolation and purification. The purity of the isolated fraction(s) can then be measured/confirmed using ACQUITY UPLC CSH columns and UPLC Technology.
1.50
AU
0.00
1.00
0.50
XSelect Preparative
Columns
10 x 10 mm 1.50
10 x 50 mm
10 x 100 mm 1.00
10 x 150 mm
10 x 250 mm 0.50
19 x 10 mm
OBD 19 x 50 mm0.00
OBD 19 x 100 mm
0.0
OBD 19 x 150 mm
OBD 19 x 250 mm
OBD 30 x 50 mm
OBD 30 x 75 mm
OBD 30 x 100 mm
OBD 30 x 150 mm
OBD 30 x 250 mm
OBD 50 x 50 mm
OBD 50 x 100 mm
OBD 50 x 150 mm
OBD 50 x 250 mm
Particle6
Size
Type 4
Guard
Column
Column
Column
Column
Guard
Column
Column
0.2
0.4
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Prep Purification
100 mg total load
0.00
AU
1.50
AU
Target Analyte
1
0.50
182
1.00
1.00
Dimension
Prep Purification
100 mg total load
4. Collect and isolate
fraction(s) of interest
1.50
Injection Volume:
Crude mix
DiphenhydramineSample
= 56.3%
purity
Concentration:
Diphenhydramine:
Oxybutynin:
Terfenadine :
1.2
1.4
1.6Sample
1.8Diluent:
2.0 min
Column Temperature:
Weak Needle Wash:
3
Strong Needle Wash:
Detection:
Sampling Rate:
Filter Time Constant:
System:
10 min
0.50
1.0
Collected Fraction
Prep Purification
100 mg total load
1.50
0.4
1
2.00
0.50
AU
Injection Volume:
Sample Concentration:
Sample Diluent:
Column Temperature:
Weak Needle Wash:
Strong Needle Wash:
Detection:
Sampling Rate:
Filter Time Constant:
System:
Preparative Conditions
Column:
Part Number:
Mobile Phase A:
Mobile Phase B:
Gradient:
0.6
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
0.8
1.0
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
Fluoro-Phenyl
Phenyl-Hexyl
1860054911
1860054981
186005414
186005427
186005415
186005428
186005416
186005429
Pooled purified fractions
186005417
186005430
Diphenhydramine
= 100% purity
1860054182
1860054312
186005420
186005433
186005421
186005434
1.2
1.4
1.6
1.8
2.0 min
186005422
186005435
186005492
186005499
186005423
186005436
186005424
186005437
186005425
186005438
186005426
186005439
186005493
186005500
186005494
186005501
186005495
186005502
186005496
186005503
186005497
186005504
1860055051
186005440
186005441
186005442
186005443
1860054442
186005446
186005447
186005448
186005506
186005520
186005450
186005451
186005452
186005507
186005508
186005509
186005510
186005511
C818
Preparative Chromatography
10 min
Description
1
2
Part No.
10 x 10 mm Cartridge Holder
19 x 10 mm Cartridge Holder
289000779
186000709
700001019
700001436
700001020
www.waters.com
PREPaRaTIVE ChROMaTOGRaPhY
XBridge Prep OBD Columns
Bridging the Performance Gap from Analytical to Prep
A Particle Designed for Purification
Preparative packing materials are routinely offered with a muc h wider
particle size distribution than the equivalent analytical particle. W hile this
approach does reduce the cost of material manufacturing, the chromatographic performance is almost certainly compromised.
XBridge Prep packing materials are fully optimized to deliver maximum
efficiency and the highest loadability at the lowest possible backpressures.
T hese attributes allow the purification chemist the flexibility to choose the
best combination of particle size and column dimension to easily purify
even the most complex of samples.
A Milestone in Chromatography
First introduced in 1999, Waters patented Hybrid Particle Technology (HPT)
surmounted significant limitations of silica-based, reversed-phase packing
materials, particularly their hydrolytic instability at high pH. In 2005, the
introduction of a second generation HPT, branded as XBridge Prep columns
marks a new milestone in chromatography.
With an order of magnitude improvement in high pH stability and a higher
level of chromatographic performance, XBridge Prep columns define the
new benchmark for LC purification.
Gradient:
Time
(min)
0.0
1.79
6.79
7.79
Profile
%A
%B
95
5
95
5
5
95
5
95
Gradient:
Time
(min)
0.0
1.79
6.79
7.79
Profile
%A
%B
95
5
95
5
5
95
5
95
5
1
Compounds
1. Labetolol (50 mg/mL)
2. Quinine (50 mg/mL)
3. Diltiazem (50 mg/mL)
4. Verapamil (100 mg/mL)
5. Amitriptyline (50 mg/mL)
www.waters.com
10
12
14
16
18
20 Min.
XBridge Prep Columns deliver the same high loading capacity and reliability expected
of our XTerra Prep products, with a significantly reduced column backpressure.
Literature
References
Waters XBridge HPLC Columns Brochure,
Literature Reference 720001255EN
BEH Technology W hite Paper,
Literature Reference 720001159EN
Optimum Bed Density (OBD) Columns:
Enabling Technology for Laboratory-Scale
Isolation and Purification W hite Paper,
Literature Reference 720001939EN
Preparative Chromatography
183
PREPARATIVE CHROMATOGRAPHY
Ligand
Density
O
OO Si
OSi
OO
OO
OSi
OOSi
Trifunctional C18
Carbon
Load
pH
Range
Pore
Diameter
Surface
Area
3.1 mol/m2
18%
1-12
130
185 m2/g
3.2 mol/m2
13%
1-12
130
185 m2/g
3.3 mol/m2
17%
2-11
130
185 m2/g
Description
3.0 mol/m2
15%
1-12
130
185 m2/g
10 x 10 mm Cartridge Holder
19 x 10 mm Cartridge Holder
289000779
186000709
n/a
unbonded
1-9
130
185 m2/g
700001019
700001436
700001020
O
OO Si
O
OO Si
OO
O Si
OO Si
Trifunctional C8
CH 3
Monofunctional Embedded
Polar Group (Shield RP18)
CH 3
O Si
CH 3
Si
CH
CH33
Si
CH3
Si
CH3
CH3
O
O
O
Polar Group
Polar Group
Polar Group
Trifunctional C6 Phenyl
Part No.
Polar Group
OO Si
OSi
OO
OO
OSi
OOSi
O
Unbonded BEH
Particle
Size
C18
10 x 10 mm
10 x 50 mm
10 x 100 mm
10 x 150 mm
10 x 250 mm
19 x 10 mm
OBD 19 x 50 mm
OBD 19 x 100 mm
OBD 19 x 150 mm
OBD 19 x 250 mm
OBD 30 x 50 mm
OBD 30 x 75 mm
OBD 30 x 100 mm
OBD 30 x 150 mm
OBD 30 x 250 mm
OBD 50 x 50 mm
OBD 50 x 100 mm
OBD 50 x 150 mm
OBD 50 x 250 mm
Guard
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
10 x 10 mm
10 x 150 mm
10 x 250 mm
19 x 10 mm
OBD 19 x 50 mm
OBD 19 x 100 mm
OBD 19 x 150 mm
OBD 19 x 250 mm
OBD 30 x 75 mm
OBD 30 x 100 mm
OBD 30 x 150 mm
OBD 30 x 250 mm
OBD 50 x 50 mm
OBD 50 x 100 mm
OBD 50 x 150 mm
OBD 50 x 250 mm
Guard
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Dimension
1
2
C8
Shield RP18
Phenyl
HILIC
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
186002972
186002973
186003255
186002974
186003256
1860029752
186002977
186002978
186002979
186004021
186002980
186002981
186002982
186003284
186004025
186003933
186003937
186003929
186004107
186002991
186003264
186003265
186003266
186003267
1860029922
186002993
186002994
186002995
186004023
186002996
186003269
186002997
186003083
186003934
186003938
186002983
186003257
186003258
186003259
186003260
1860029842
186002985
186002986
186002987
186004022
186002988
186003262
186002989
186002990
186003935
186003939
186003354
186003271
186003272
186003273
186003274
1860033552
186003356
186003357
186003358
186004024
186003277
186003278
186003279
186003276
186003936
186003940
1860047201
186004721
186004722
1860047232
186004724
186004725
186004726
186004730
186004727
186004728
186004729
186004731
186004732
186004733
186004734
186004735
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
1860038893
186003890
186003891
1860038924
186003893
186003901
186003894
186003895
186004711
186003930
186003896
186003897
186003898
186003902
186003899
186003900
1860040033
186004004
186004005
1860040064
186004007
186004008
186004009
186004010
186004011
186004012
186004013
186004014
186004015
186004016
1860039883
186003989
186003990
1860039914
186003992
186003993
186003994
186003995
186003996
186003997
186003998
186003999
186004001
186004002
For Purification
of Peptides, See
Pages 226-235
184
Preparative Chromatography
www.waters.com
PREPaRaTIVE ChROMaTOGRaPhY
SunFire Prep OBD Columns
Bridging the Performance Gap from Analytical to Prep
Isolation of Nimodipine and Its Impurities
Mobile Phase A:
Mobile Phase B:
Analyte:
Detection:
Instruments:
Impurity levels:
Flow Rate:
Gradient:
Nimodipine
100
SunFire C18
4.6 x 100 mm, 5 m
Part Number: 186002558
Total Mass Load: 6 mg
%
100
-1
%
SunFire columns set the standard for the state-of-the-art bonded C18
and C 8 silica HPLC columns. Benefiting from years of researc h and
product development, SunFire columns represent the best in particle
and bonding expertise and deliver the industry-leading level of
c hromatographic performance.
Flow Rate:
Injection Volume:
Column:
Detection:
Instrument:
SunFire C18
Rs: 2.77
W1/2: 0.17
%
100
1
SunFire Prep OBD C18
19 x 100 mm, 5 m
Part Number: 186002567
Total Mass Load: 102 mg
-1
%
Kromasil C18
250 g
Rs: 1.19
W1/2: 0.53
mg/mL
25
50
50
125
mg/mL
20
40
40
100
mg/mL
10
20
20
50
mg/mL
5
10
10
25
Compounds:
1. Ketoconazole
2. Econazole
3. Miconazole
3
4
2
8
10
10
1.4 mL/min
Time
Profile
(min)
%A
0.00
70
1.00
70
11.00 10
12.00 10
12.10 70
16.00 70
200 L
23.9 mL/min
Time
Profile
(min)
%A
0.00
70
1.97
70
11.97 10
12.97 10
13.10 70
12
14 Min.
16.00 70
Inj. Vol.:
3.4 mL
%B
30
30
90
90
30
30
Flow Rate:
Gradient:
12
%B
30
30
90
90
30
30
14 Min.
SunFire Prep silica columns allow you to use normal-phase protocols. T his
gives you a significant selectivity alternative to reversed-phase preparative
c hromatography on bonded silicas. Analytes that are highly hydrophilic
or that are insoluble/unstable in water are ideal candidates for normalphase purification. Unlike the more viscous, higher-boiling aqueous-organic
mobile phases used in reversed phase, typical normal-phase eluents are
lower-viscosity, lower-boiling organic solvents. As a consequence, SunFire
Silica separations yield several key benefits:
Lower backpressure
Literature
References
500 g
Rs: 1.01
W1/2: 0.64
Rs: 1.92
W1/2: 0.26
1000 g
Rs: 0.81
W1/2: 0.90
1250 g
Rs: 0.78
W1/2: 0.91
www.waters.com
Inj. Vol.:
Rs: 2.31
W1/2: 0.21
Rs: 1.87
W1/2: 0.27
% by Area
3.1
0.6
6.5
4.9
Sample Concentration:
As indicated in the table, dissolved in DMSO
Analyte
Ketoconazole
Econazole
Miconazole
Total Concentration
2
-1
Loadability by Design
Mobile Phase A:
Mobile Phase B:
Gradient:
100
-1
Peak
1
2
3
4
Preparative Chromatography
185
PREPARATIVE CHROMATOGRAPHY
O
O
HO
O
O
O
O
O
HO
OH
O
Conditons
Column:
Part Number:
Mobile Phase:
Analytes:
1. Propyl p-phenylacetobenzoate
2. Methyl p-phenylacetobenzoate
3. Propyl paraben
4. Methyl paraben
Flow Rate:
Total Load:
O
O
O
OH
HO
Phenylacetic acid
Propyl paraben
O
O
Propyl p-phenylacetobenzoate
DCC/DMAP in THF
Methyl p-phenylacetobenzoate
Methyl paraben
O
O
HO
10
12 Min
Dimension
10 x 50 mm
10 x 100 mm
10 x 150 mm
10 x 250 mm
OBD 19 x 50 mm
OBD 19 x 100 mm
OBD 19 x 150 mm
OBD 19 x 250 mm
OBD 30 x 50 mm
OBD 30 x 75 mm
OBD 30 x 100 mm
OBD 30 x 150 mm
OBD 30 x 250 mm
OBD 50 x 50 mm
OBD 50 x 100 mm
OBD 50 x 150 mm
OBD 50 x 250 mm
Particle
Size
C18
C8
Silica
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
186002561
186002562
186002563
186002564
186002566
186002567
186002568
186004027
186002570
186002571
186002572
186002797
186003969
186002867
186002869
186003941
186003970
186002746
186002747
186002748
186002749
186002751
186002752
186002753
186004028
186002755
186002756
186002757
186002795
186002868
186002870
186003425
186003426
186003427
186003428
186003431
186003432
186003433
186004029
186003435
186003436
186003437
186003438
186003439
186003440
Dimension
Particle
Size
C18
C8
Silica
10 x 50 mm
10 x 150 mm
10 x 250 mm
OBD 19 x 50 mm
OBD 19 x 150 mm
OBD 19 x 250 mm
OBD 30 x 50 mm
OBD 30 x 100 mm
OBD 30 x 150 mm
OBD 30 x 250 mm
OBD 50 x 50 mm
OBD 50 x 100 mm
OBD 50 x 150 mm
OBD 50 x 250 mm
OBD 100 x 250 mm
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
186003840
186002664
186002665
186002667
186002668
186002669
186003854
186003971
186002670
186002671
186002871
186003972
186002672
186002673
186003928
186003841
186002759
186002760
186002762
186002763
186002764
186003853
186002765
186002766
186002872
186002767
186002768
Custom
186003442
186003443
186003445
186003446
186003447
186003855
186003448
186003449
186003450
186003451
186003452
Dimension
Particle
Size
C18
10 x 10 mm
19 x 10 mm
10 x 10 mm
19 x 10 mm
5 m
5 m
10 m
10 m
186002565
1860025692
1860026631
1860026662
C8
1
Silica
186002750
1860027542
1860027581
1860027612
1
1860034291
1860034342
1860034411
1860034442
Description
1
2
Dimension
Particle
Size
C18
C8
Silica
4.6 x 150 mm
4.6 x 250 mm
4.6 x 150 mm
4.6 x 250 mm
5 m
5 m
10 m
10 m
186003390
186003391
Custom
Custom
186003453
186003454
186003467
186003468
186
Preparative Chromatography
Part No.
186000708
289000779
186000709
700001019
700001436
700001020
www.waters.com
PREPaRaTIVE ChROMaTOGRaPhY
Atlantis Prep OBD Columns
Bridging the Performance Gap for Polar Compound Retention
Beta Blockers
Atlantis T3
4.6 x 100 mm, 5 m
Part Number: 186003746
3.0
2
AU
2.0
Flow Rate:
Inj. Vol.:
V 0 = 1.5 min
1.0
1.0 mL/min
15 L (6.6 mg total mass load)
0.0
3
3.0
2
AU
2.0
1.0
Flow Rate:
Inj. Vol.:
V 0 = 1.5 min
17.06 mL/min
256 L (112.6 mg total mass load)
0.0
3
3.0
2
AU
2.0
First introduced in 2002, the Atlantis product range has rapidly gained
acceptance as the industry standard for polar compound retention and
purification. Available for both reversed-phase and HILIC modes of
c hromatography, these phases exhibit significantly increase retention
c haracteristics as compared to conventional C18 or embedded polar
group column c hemistries.
Atlantis dC18
T he first in the family to be introduced, Atlantis dC18 is a reversedphase packing bonded with a difunctional C18 ligand to an intermediate
ligand density (1.6 moles/m2) and fully end-capped. For maximum
column lifetime, the dC18 phase is recommended for mobile-phase pH
values of 2.56.
Atlantis T3
An evolution of the dC18 product, Atlantis T3 columns retain all of the
benefits of its predecessor but exhibit significantly improved stability
in low-pH mobile phases as well as improved peak shape for amine
containing bases at pH 7. T3 bonding utilizes a trifunctional C18 ligand,
(1.6 moles/m2), and a proprietary end-capping process whic h has been
proven to be muc h more effective than the traditional trimethyl silane
(TMS) end-capping.
V 0 = 1.5
1.0
min
Flow Rate:
Inj. Vol.:
42.53 mL/min
638 L (280.7 mg total mass load)
0.0
0
Profile
%A
100
100
82
28
28
0
0
100
100
Profile
%A
100
100
82
28
28
0
0
100
100
%B
0
0
18
72
72
100
100
0
0
10
12 Min.
Compounds
1. Atenolol
2. Metoprolol
3. Propranolol
40 mg/mL in DMSO
200 mg/mL in DMSO
200 mg/mL in DMSO
Mobile Phase A:
Mobile Phase B:
Detection:
Instrument:
Temperature:
%B
0
0
18
72
72
100
100
0
0
Literature
References
www.waters.com
Preparative Chromatography
187
PREPARATIVE CHROMATOGRAPHY
HILIC is a variation of normal-phase chromatography where a polar stationary phase is used with a mobile phase which contains a high concentration of
organic (non-polar) solvent and a low concentration of aqueous (polar)
solvent. In HILIC, the organic portion of the mobile phase is the weak
solvent, the aqueous portion the strong solvent and the compounds elute in
the order of increasing hydrophilicity. Due to the high organic, low viscosity
mobile phases that are employed, HILIC provides low column backpressure. Additionally, the collected fractions of interest are inherently high in
organic, enabling faster solvent evaporation compared to reversed phase.
Columns:
Mobile Phase A:
Mobile Phase B:
Gradients:
Flow Rates:
Sample Concentration:
Injection Volumes:
Weak Wash Solvent:
Temperature:
Detection:
Instrument:
Gradient:
Time
Profile
(min)
%A
%B
0.0
0
100
5.0
10
90
6.0
10
90
6.1
0
100
10.0
0
100
Injection Volume: 60 L
Total Loading:
4500 ng
2
3
Gradient:
Time
Profile
(min)
%A
%B
0.0
0
100
0.74
0
100
5.74
10
90
6.74
10
90
6.84
0
100
10.74 0
100
Injection Volume: 1020 L
Total Loading:
76.5 g
2
3
1
Type
Particle
Size
T3
10 x 10 mm
10 x 50 mm
10 x 100 mm
10 x 150 mm
10 x 250 mm
19 x 10 mm
OBD 19 x 50 mm
OBD 19 x 100 mm
OBD 19 x 150 mm
OBD 19 x 250 mm
OBD 30 x 50 mm
OBD 30 x 75 mm
OBD 30 x 100 mm
OBD 30 x 150 mm
OBD 50 x 50 mm
OBD 50 x 100 mm
OBD 50 x 150 mm
Guard
Column
Column
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
10 x 10 mm
10 x 150 mm
10 x 250 mm
19 x 10 mm
OBD 19 x 50 mm
OBD 19 x 150 mm
OBD 19 x 250 mm
OBD 30 x 75 mm
OBD 30 x 150 mm
OBD 30 x 250 mm
OBD 50 x 50 mm
OBD 50 x 100 mm
OBD 50 x 150 mm
OBD 50 x 250 mm
Guard
Column
Column
Guard
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
Dimension
188
Preparative Chromatography
HILIC
dC18
186003695
186003691
186003692
186003693
186003694
1860036992
186003696
186003697
186003698
186004026
186003700
186003701
186003702
186003703
186004080
186004081
186004082
1860039562
186003957
186003958
186003959
186003960
186003961
186003962
1860023001
186002298
186002299
1860013612
186001365
186001367
186002800
186004030
186001373
186002455
186001375
186002801
1860037061
186003704
186003705
1860037102
186003707
186003708
186003709
186004712
186003711
186003712
186004083
186004084
186004085
186004086
1860024521
186002453
186002454
1860013632
186001369
186001371
186002417
186002418
Compounds
1. Uracil
2. 5-Fluorocytosine
3. Cytosine
4 Min.
1
2
Part No.
10 x 10 mm Cartridge Holder
19 x 10 mm Cartridge Holder
289000779
186000709
700001019
700001436
700001020
www.waters.com
PREPARATIVE CHROMATOGRAPHY
XTerra Prep OBD Columns
Bridging the Performance Gap from Analytical to Prep
Particle
Size
MS C18
MS C8
RP18
RP8
7.8 x 10 mm
7.8 x 50 mm
7.8 x 100 mm
7.8 x 150 mm
10 x 10 mm
10 x 30 mm
10 x 50 mm
10 x 100 mm
10 x 150 mm
19 x 10 mm
OBD 19 x 50 mm
OBD 19 x 100 mm
OBD 19 x 150 mm
OBD 30 x 50 mm
OBD 30 x 75 mm
OBD 30 x 100 mm
OBD 50 x 50 mm
OBD 50 x 100 mm
Cartridge
Column
Column
Column
Cartridge
Column
Column
Column
Column
Cartridge
Column
Column
Column
Column
Column
Column
Column
Column
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
1860011686
186001152
186001156
186001475
1860010017
186001010
186001014
186001018
186001479
1860011048
186001930
186001934
186002379
186001938
186002387
186001942
186002218
186002222
1860011696
186001153
186001157
186001476
1860010047
186001011
186001015
186001019
186001480
1860011058
186001931
186001935
186002380
186001939
186002388
186001943
186002219
186002223
1860011706
186001154
186001158
186001477
1860010067
186001012
186001016
186001020
186001481
1860011068
186001932
186001936
186002381
186001940
186002389
186001944
186002220
186002224
1860011716
186001155
186001159
186001478
1860010087
186001013
186001017
186001021
186001482
1860011078
186001933
186001937
186002382
186001941
186002390
186001945
186002221
186002225
7.8 x 10 mm
7.8 x 150 mm
7.8 x 300 mm
10 x 10 mm
10 x 150 mm
10 x 250 mm
10 x 300 mm
19 x 10 mm
OBD 19 x 50 mm
OBD 19 x 150 mm
OBD 19 x 250 mm
OBD 19 x 300 mm
OBD 30 x 150 mm
OBD 30 x 250 mm
OBD 30 x 300 mm
OBD 50 x 50 mm
OBD 50 x 150 mm
OBD 50 x 250 mm
Cartridge
Column
Column
Cartridge
Column
Column
Column
Cartridge
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
1860011726
186001160
186001164
1860010027
186001022
186001026
186001030
1860010348
186002254
186002255
186002259
186002263
186002267
186002271
186002275
186002279
186002843
186002847
1860011736
186001161
186001165
1860010057
186001023
186001027
186001031
1860010358
186002256
186002260
186002264
186002268
186002272
186002276
186002280
186002844
186002848
1860011746
186001162
186001166
1860010077
186001024
186001028
186001032
1860010368
186002257
186002261
186002265
186002269
186002273
186002277
186002281
186002845
186002849
1860011756
186001163
186001167
1860010097
186001025
186001029
186001033
1860010378
186002258
186002262
186002266
186002270
186002274
186002278
186002282
186002846
186002850
Dimension
XTerra Chemistries
XTerra MS
XTerra MS C18 and MS C8 columns were designed to be
compatible with Mass Spectrometry applications and
provide sharp peaks, good sensitivity, and large peak
capacities. T hese chemistries exhibit peak shape usually
only ac hievable with columns employing embedded
polar group tec hnology (see XTerra RP columns). T he
tri-functional bonding chemistries of XTerra MS columns
deliver the longest lifetimes over the widest pH range
(pH 1-12) and provide maximum throughput with
excellent resolution and ultra-low bleed.
XTerra RP
XTerra R P18 and R P8 columns combine Hybrid
Particle Tec hnology wit h S hield Tec hnology by
incorporating an embedded polar group to deliver
the best possible peak shape. T hese general purpose
columns allow fast method development by offering unique selectivity. Like all XTerra columns, the
shielded XTerra RP columns exhibit excellent water
wettability, even in 100% aqueous mobile phases.
T he combination of a greatly widened pH range
for selectivity choices and the additional selectivity
options of two ligands are often the only tools
necessary for successful method development.
www.waters.com
Part No.
10 x 10 mm Cartridge Holder
19 x 10 mm Cartridge Holder
289000779
186000709
700001019
700001436
700001020
Preparative Chromatography
189
PREPARATIVE CHROMATOGRAPHY
Symmetry Columns
SymmetryPrep Columns for Purification and Isolation
Dimension
SymmetryShield Columns
C18
C8
7.8 x 10 mm
7.8 x 50 mm
7.8 x 100 mm
19 x 10 mm
19 x 50 mm
19 x 100 mm
30 x 50 mm
30 x 100 mm
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
186000711
186000208
186000209
1860007151
186000210
186000211
186000235
186000236
186000712
186000214
186000215
1860007161
186000216
186000229
186000237
186000238
7.8 x 10 mm
7.8 x 150 mm
7.8 x 300 mm
19 x 10 mm
19 x 150 mm
19 x 300 mm
7 m
7 m
7 m
7 m
7 m
7 m
186000713
WAT066288
WAT066235
1860007171
WAT066240
WAT066245
186000714
WAT066285
WAT066225
1860007181
WAT066228
WAT066230
SymmetryPrep Columns
SymmetryPrep 7 m columns are manufactured for t he c hemist w ho
needs to purify new chemical entities or isolate impurities or degradants.
T he SymmetryPrep 7 m products are a family of high-capacity, highefficiency, high-performance liquid chromatography (HPLC) columns that
have identical selectivity to t he Symmetry 3.5 and 5 m particle
size analytical columns. T his guarantees direct method scale-up with no
further development. SymmetryPrep columns deliver the advantages of
unmatched peak shape and resolution using simple volatile mobile-phase
buffers such as TFA and ammonium acetate. All of these features result in a
tremendous time reduction in your research and development programs.
Particle Size
Particle Size
RP18
19 x 10 mm
19 x 50 mm
19 x 100 mm
19 x 150 mm
5 m
5 m
5 m
5 m
1860018351
186001836
186001837
186001838
1860018411
186001842
186001843
186001844
RP8
19 x 150 mm
19 x 300 mm
7 m
7 m
186001839
186001840
186001845
186001846
Particle Size
5 m
5 m
5 m
5 m
C18
1860018471
186001848
186001849
186001850
Symmetry300 Columns
Symmetry300, reversed-phase columns are designed specifically for high
recoveries of peptides and proteins. Symmetry300 columns are the best
solution for validation compliance regarding identity, purity, and stability.
190
Preparative Chromatography
Part No.
186000708
289000779
186000709
700001019
700001436
700001020
www.waters.com
PREPaRaTIVE ChROMaTOGRaPhY
Waters Spherisorb Columns
Waters Spherisorb columns are the most widely referenced HPLC columns
in scientific literature. T here are over 2,000 abstracts published on
Spherisorb columns, providing a tremendous range of validated methods
and applications to assist you in your method development process.
10 x 250 mm
20 x 250 mm
S10 ODS2
S10 ODS1
S10 C8
S10 C6
S10 C1
S10 NH2
S10 P
S10 CN
S10 W
PSS832585
PSS830785
PSS832885
PSS833285
PSS833085
PSS833685
PSS833885
PSS833585
PSS830285
PSS832595
PSS830795
PSS832895
PSS833295
PSS833095
PSS833695
PSS833895
PSS833595
PSS830295
PSS833985
PSS837685
PSS833995
PSS837695
Dimension
Part No.
3.9 x 300 mm
7.8 x 300 mm
19 x 300 mm
3.9 x 300 mm
7.8 x 300 mm
19 x 300 mm
WAT038500
WAT025820
WAT025822
WAT038501
WAT025821
WAT025823
Dimension
Part No.
3.9 x 300 mm
7.8 x 300 mm
19 x 150 mm
19 x 300 mm
3.9 x 300 mm
7.8 x 300 mm
19 x 300 mm
3.9 x 300 mm
7.8 x 300 mm
7.8 x 300 mm
3.9 x 300 mm
7.8 x 300 mm
3.9 x 300 mm
7.8 x 300 mm
19 x 150 mm
19 x 300 mm
3.9 x 300 mm
19 x 300 mm
WAT027324
WAT084176
WAT088500
WAT025828
WAT025875
WAT025832
WAT025834
WAT027198
WAT084179
WAT084177
WAT084040
WAT084178
WAT027477
WAT084175
WAT091648
WAT025829
WAT025874
WAT025835
Ion Exchange
S10 SAX
S10 SCX
Description
Particle
Size
Pore
Size
6 m
60
6 m
60
Bondapak/Porasil Columns
T he popular Bondapak C18 chemistry and Porasil silica packing materials
are available in three different particle sizes, 10, 15-20, and 37-55 m,
providing easy transfer of c hromatography met hodology from one
particle size to another. You can utilize your existing 10 m Bondapak
or Porasil c hromatography as a starting point for scale-up or, if you are
developing a new purification protocol, investigate whic h preparative
particle size provides the best balance between resolution, throughput
and cost.
T he preparative Bondapak HC18 HA (high-carbon load, high-activity silica) is
a highly carbon-loaded packing that provides different selectivity over the
standard Bondapak packing materials. T he higher carbon load on the silica
surface typically results in a higher loading capability. Bondapak HC18 HA
is available in a 37-55 m particle size.
T he Prep family of packing materials is based on a 55-105 m Porasil
silica and provides a cost effective means for scale-up of preparative
processes. T he selectivity of Prep silica is identical to 10, 15-20, and
37-55 m Porasil while Prep C18 has a slightly different selectivity than
Bondapak packings.
www.waters.com
Description
Particle
Size
Pore
Size
Bondapak C18
10 m
125
Bondapak C18
15-20 m
125
Bondapak Phenyl
10 m
125
Bondapak CN
Bondapak NH2
10 m
10 m
125
125
Porasil
10 m
125
Porasil
15-20 m
125
Preparative Chromatography
191
PREPaRaTIVE ChROMaTOGRaPhY
Delta-Pak Packing
Delta-Pak packing materials are ideal for the separation of peptides,
proteins, and natural products. T he isolation and purification of a peptide
is usually a multi-step procedure with the fractions from the first run
being re-chromatographed on the same preparative column to obtain pure
product. Delta-Pak packing materials are based on a highly stable, bonded,
endcapped 5 and 15 m packing. T he 5 m packing is available in analytical scale dimensions for preliminary preparative chromatographic studies,
peptide mapping, and fraction purity assays. T he chemistry characteristics
of the packing materials are independent of the particle size.
Delta-Pak Columns
Description
Particle
Size
Pore
Size
Dimension
Part No.
Delta-Pak C18
15 m
Delta-Pak C4
15 m
100
300
100
300
100
300
100
300
100
100
300
100
300
100
300
100
300
3.9 x 300 mm
3.9 x 300 mm
7.8 x 300 mm
7.8 x 300 mm
19 x 300 mm
19 x 300 mm
30 x 300 mm
30 x 300 mm
50 x 300 mm
3.9 x 300 mm
3.9 x 300 mm
7.8 x 300 mm
7.8 x 300 mm
19 x 300 mm
19 x 300 mm
30 x 300 mm
30 x 300 mm
WAT011797
WAT011802
WAT011798
WAT011803
WAT011799
WAT011804
WAT011800
WAT011805
WAT011801
WAT011807
WAT011812
WAT011808
WAT011813
WAT011809
WAT011814
WAT011810
WAT011815
Particle
Size
Pore
Size
Porasil Silica
15-20 m
125
Porasil Silica
37-55 m
125
Prep Silica
55-105 m
125
Description
192
Dimension
Part No.
100 g
1 kg
5 kg
100 g
1 kg
5 kg
100 g
1 kg
5 kg
WAT020731
WAT020732
WAT020733
WAT020721
WAT020722
WAT020723
WAT020587
WAT010004
WAT020588
Preparative Chromatography
Particle
Size
Pore
Size
Bondapak C18
15-20 m
125
Bondapak C18
37-55 m
125
Bondapak HC18 HA
37-55 m
125
Prep C18
55-105 m
125
Description
Dimension
Part No.
100 g
1 kg
5 kg
100 g
1 kg
5 kg
100 g
1 kg
5 kg
100 g
1 kg
5 kg
WAT020739
WAT020740
WAT020741
WAT030632
WAT030633
WAT030634
WAT035672
WAT035674
WAT035676
WAT020594
WAT010001
WAT020595
www.waters.com
PREPARATIVE CHROMATOGRAPHY
Radial Compression Modules
We carry a complete inventory of accessories and spare parts for the Waters
patented Radial Compression Modules for use with the 5 mm and 8 mm i.d.
Radial-Pak column segments, the 25 mm and 40 mm i.d. PrepLC column
segments, and the 47 mm and 57 mm i.d. PrepPak cartridges.
Particle Size
Pore Size
Dimension
Part No.
5 x 100 mm
8 x 100 mm
5 x 100 mm
8 x 100 mm
5 x 100 mm
8 x 100 mm
5 x 100 mm
8 x 100 mm
5 x 100 mm
8 x 100 mm
WAT080100
WAT086342
WAT035890
WAT035884
WAT010657
WAT010658
WAT010224
WAT010223
WAT010986
WAT010987
8 x 100 mm
8 x 100 mm
WAT025843
WAT025844
8 x10 Holder
8 x 10 Holder for 8 x 100 mm and
5 x 100 mm Radial-Pak column segments
Description
Part No.
8 x 10 Holder
8 x 10 Extension Kit (includes 1 extension tube, union, o-rings)
Cartridge Union
WAT082887
WAT038846
WAT038849
Nova-Pak C18
4 m
60
Nova-Pak C8
4 m
60
Nova-Pak Phenyl
4 m
60
Nova-Pak CN HP
4 m
60
Nova-Pak Silica
4 m
60
PrepLC 25 mm Module
6 m
6 m
60
60
Description
Part No.
PrepLC 25 mm Module
PrepLC 25 mm Extension Kit (includes 1 extension tube, union, o-rings)
Extension Tube
WAT015814
WAT022180
WAT019311
6 m
6 m
60
60
25 x 100 mm WAT038510
25 x 100 mm WAT038511
6 m
6 m
60
60
25 x 10 mm
25 x 10 mm
WAT038528
WAT038530
PrepLC Assemblies
PrepLC Assembly for
40 x 100 mm and
25 x 100 mm
column segments
PrepLC
Assembly*
40 x 100 mm
www.waters.com
40 x 100 mm WAT037704
40 x 100 mm WAT037708
6 m
6 m
60
60
40 x 10 mm
40 x 10 mm
WAT037854
WAT037857
PrepLC 40 mm Assembly
(includes PrepLC Universal Base and PrepLC 40 mm Chamber)
PrepLC Universal Base
PrepLC 40 mm Chamber (includes o-rings, spacer and union)
PrepLC 40 mm Extension Kit (includes extension tube, union, o-rings)
PrepLC 25 mm Chamber (includes o-rings, spacer and union)
PrepLC 25 mm Extension Kit (includes 1 extension tube, union, o-rings)
PrepLC Scale-up Kit with capability for 40 mm or 25 x 300 length
includes: 1 - PrepLC Universal Base
2 - PrepLC Chambers (one each of 40 mm and 25 mm)
2 - PrepLC 25 mm Extension Kits
2 - PrepLC 40 mm Extension Kits
60
60
PrepLC
Assembly*
25 x 100 mm
Description
6 m
6 m
Part No.
WAT022441
WAT027577
WAT027578
WAT022365
WAT033994
WAT022180
WAT022440
Particle Size
Pore Size
Dimension
Part No.
Resolve C18
Resolve C18
5 m
10 m
90
90
Resolve C8
10 m
90
Resolve CN
10 m
90
Resolve Silica
Resolve Silica
5 m
10 m
90
90
8 x 100 mm
5 x 100 mm
8 x 100 mm
5 x 100 mm
8 x 100 mm
5 x 100 mm
8 x 100 mm
8 x 100 mm
5 x 100 mm
8 x 100 mm
WAT084624
WAT084620
WAT084720
WAT085672
WAT085670
WAT084626
WAT084636
WAT084634
WAT084630
WAT084730
Preparative Chromatography
193
PREPARATIVE CHROMATOGRAPHY
Particle Size
Pore Size
Part No.
10 m
10 m
10 m
10 m
15-20 m
37-55 m
10 m
15-20 m
Bondapak HC18 HA
Porasil
Porasil
37-55 m
10 m
15-20 m
Particle Size
125
125
125
125
125
125
8 x 100 mm
8 x 100 mm
8 x 100 mm
8 x 100 mm
8 x 100 mm
8 x 100 mm
WAT085721
WAT085724
WAT085722
WAT085720
WAT025841
WAT036561
125
125
300
125
125
125
25 x 100 mm
25 x 100 mm
25 x 100 mm
25 x 100 mm
25 x 100 mm
25 x 100 mm
WAT038505
WAT038503
WAT038575
WAT056970
WAT038504
WAT038502
25 x 10 mm
25 x 10 mm
25 x 10 mm
25 x 10 mm
WAT038518
WAT038514
WAT038516
WAT038512
40 x 100 mm
40 x 100 mm
40 x 100 mm
40 x 100 mm
40 x 100 mm
WAT037684
WAT037676
WAT037712
WAT037680
WAT037672
40 x 10 mm
40 x 10 mm
40 x 10 mm
40 x 10 mm
WAT037839
WAT037833
WAT037836
WAT037830
Delta-Pak C18
15 m
Delta-Pak C4
15 m
10 m
15-20 m
10 m
15-20 m
Delta-Pak C18
15 m
Delta-Pak C4
15 m
10 m
15-20 m
Porasil
Porasil
10 m
15-20 m
Part No.
100
300
100
8 x 100 mm
8 x 100 mm
8 x 100 mm
WAT025846
WAT025845
WAT025848
100
300
100
300
25 x 100 mm
25 x 100 mm
25 x 100 mm
25 x 100 mm
WAT038506
WAT038507
WAT038508
WAT038509
100
300
100
300
25 x 10 mm
25 x 10 mm
25 x 10 mm
25 x 10 mm
WAT038520
WAT038522
WAT038524
WAT038526
100
300
100
300
40 x 100 mm
40 x 100 mm
40 x 100 mm
40 x 100 mm
WAT037688
WAT037692
WAT037696
WAT037700
40 x 10 mm
40 x 10 mm
40 x 10 mm
WAT037842
WAT037845
WAT037851
15 m
Delta-Pak C4
15 m
125
125
125
125
15 m
Delta-Pak C4
15 m
Dimension
Pore Size
Column
125
125
300
125
125
15 m
Delta-Pak C4
15 m
100
300
300
10 m
15-20 m
10 m
15-20 m
125
125
125
125
PrepPak Cartridges
Requires PrepPak 1000 Module
(Part No. WAT089592)
Column
Particle Size
Pore Size
Dimension
Part No.
Bondapak C18
Bondapak C18
Bondapak C18
Bondapak HC18 HA
Bondapak NH2
Delta-Pak C18
15-20 m
15-20 m
37-55 m
37-55 m
55-105 m
15 m
125
300
125
125
125
100
300
100
300
125
125
47 x 300 mm
47 x 300 mm
47 x 300 mm
47 x 300 mm
47 x 300 mm
47 x 300 mm
47 x 300 mm
47 x 300 mm
47 x 300 mm
47 x 300 mm
47 x 300 mm
WAT091784
WAT038571
WAT025873
WAT038570
WAT091631
WAT015401
WAT010988
WAT011633
WAT011669
WAT025876
WAT025852
WAT025878
WAT025853
WAT025877
WAT089592
Delta-Pak C4
15 m
Prep C18
55-105 m
Porasil Silica (single)
15-20 m
(10/case)
Porasil Silica (single)
37-55 m
125
47 x 300 mm
(10/case)
PrepPak 1000 Module for 47 x 300 mm PrepPak Cartridges
194
Preparative Chromatography
www.waters.com
PREPARATIVE CHROMATOGRAPHY
Accessories and Spare Parts for Waters Radial
Compression Cartridge Holders and Modules
We carry a complete inventory of accessories and spare parts for
Waters patented Radial Compression Modules for use with the 5 mm
and 8 mm i.d. x 100 mm long Radial-Pak column segments, 25 mm
and 40 mm i.d. x 100 mm long PrepLC column segments, and 47 mm or
57 mm i.d. x 300 mm long PrepPak cartridges.
Part No.
WAT015814
WAT022180
WAT019311
WAT015831
WAT015833
WAT015848
WAT015834
WAT005129
WAT015854
WAT015860
WAT005137
Description
Part No.
WAT082853
WAT022281
WAT005129
WAT027509
WAT025604
WAT033996
WAT033997
WAT022453
WAT022454
WAT022280
WAT027519
WAT022299
WAT022297
WAT027518
WAT022283
WAT015835
WAT022454
WAT015860
WAT015859
WAT022276
WAT015831
WAT015833
WAT015848
WAT022298
WAT022297
WAT015834
WAT022282
WAT015835
WAT022400
www.waters.com
Preparative Chromatography
195
TM
Si
2-Ethylpyridine Ligand
196
Compounds:
1. Caffeine
2. Amitriptyline
3. Thymine
4. Cytosine
5. Sulfamethoxazole
6. Sulfamethizole
300
250
Absorbance (mAU)
200
150
100
50
0
-50
0
Mobile Phase:
5-40% methanol with
no additive
Mobile Phase:
5-40% methanol with
0.2% diethylamine (DEA)
10 min
Mobile Phase:
5-40% methanol with
20 mM ammonium acetate
www.waters.com
ORDERING INFORMATION
Part Number:
Gradient Separation:
Mobile Phase:
Flow Rate:
Sample Concentration:
Wavelength:
System Backpressure:
Compounds:
80 mg/mL
220 nm
120 bar
1. Coumarin
2. Flavone
3. Ibuprofen
4. Carbamazepine
0.25
0.50
0.75
1.00
1.25
1.50
1.75
2.00
2.25
2.50
2.75
3.00 min
Chemistry
2-Ethylpyridine
Silica
Format
Column
Column
Column
Column
Column
Column
Column
Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
Particle
Shape
Spherical
Spherical
Dimension
4.6 x 50 mm
4.6 x 100 mm
4.6 x 150 mm
4.6 x 250 mm
10 x 50 mm
10 x 100 mm
10 x 150 mm
10 x 250 mm
19 x 50 mm
19 x 100 mm
19 x 150 mm
19 x 250 mm
30 x 50 mm
30 x 75 mm
30 x 100 mm
30 x 150 mm
30 x 250 mm
50 x 50 mm
50 x 100 mm
50 x 150 mm
50 x 250 mm
Pore Size
()
100
100
Surface
Area (m2/g)
340
340
Pore Vol.
(cc/g)
0.90
0.90
Carbon
Load (%)
8
n/a
Endcapped
no
n/a
Particle
Size
Viridis SFC
2-Ethylpyridine
Viridis SFC
Silica
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
186004935
186004936
186004937
186004938
186004939
186004940
186004941
186004942
186004943
186004944
186004945
186004946
186004947
186004948
186004949
186004950
186004951
186004952
186004953
186004954
186004955
186004908
186004909
186004910
186004911
186004912
186004913
186004914
186004915
186004916
186004917
186004918
186004919
186004920
186004921
186004922
186004923
186004924
186004925
186004926
186004927
186004928
Compounds:
1. Flavone
2. Ibuprofen
3. Carbamazepine
4.2 min
5-15% methanol with no additive
85 g/min at 40 C
220 nm
120 bar
1
6.0e-1
4.0e-1
2.0e-1
Literature
References
Viridis SFC Columns Brochure,
Literature Reference 720003326EN
Basic Principles for Large-Scale Supercritical
Fluid Chromatography Application Note,
Literature Reference 720003711EN
Optimum Bed Density (OBD) Columns:
Enabling Technology for Laboratory-Scale
Isolation and Purification W hite Paper,
Literature Reference 720001939EN
Interactive Waters Reversed-Phase
Column Selectivity Chart,
www.waters.com/selectivitychart
0.0
6.0e-1
2
3
4.0e-1
2.0e-1
0.0
0.00
0.50
1.00
1.50
2.00
2.50
3.00
3.50
4.00
4.50 min
Using the same SFC gradient conditions, different selectivity can be achieved by using a Viridis SFC 2-Ethylpyridine
column or Viridis SFC silica column. The retention time for ibuprofen was reduced and the carbamazepine retention
time was increased by switching SFC column chemistries, with little change in the retention of flavone.
www.waters.com
197
198
199
200
AccQTag
Glycan Analysis
Glycoprotein analysis can involve identifying complex N and 0-Linked structures composed of
frequently similar and repeating sugar moieties. Hydrophilic-interaction liquid chromatography
(HILIC) with fluorescence detection is a well-recognized and reliable technique that effectively
separates and quantitates isolated glycans after their derivatization with fluorescent labels.
Waters ACQUITY UPLC BEH Glycan column was specifically developed and QC tested to provide
superior UPLC component resolution in less time for a range of glycan structures. W hen coupled
to Waters ACQUITY UPLC system, our column chemistry is ideally suited to help users get the
correct answers faster.
www.waters.com
201
best retention and solubility for peptides. It also gives the best peak
shape since the peptide basic groups are shielded from secondary interactions with the surface of the packing material.
TFA, however, reduces the sensitivity of MS detection of peptides. W hile
usable signal is usually obtained by using lower concentrations of TFA,
many investigators prefer to use formic acid as the mobile phase modifier.
In this case, the column surface has a larger effect on the separation.
Retention is often muc h less, and many of the peptide peaks will be
asymmetrical and tailing. It is common practice to choose columns designated as no TFA or low TFA for these separations. T he BioSuite PA-A
column described below was developed for this purpose. More recently the
development of Ethylene-Bridged Hybrid (BEH Technology) packing materials provided an improved option. T he ethane bridges reduce the surface
concentration of silanol groups so that there is little secondary interaction
between peptides and the base particle. Packing materials based on these
particles can be used for peptide separations in either T FA or formic
acid containing mobile phases without significant loss of retention or
band-broadening. T he BEH Tec hnology particles for the basis of the
Peptide Separation Technology family of columns described below.
With the Peptide Separation Technology columns, the minimal secondary
interactions create the greatest number of options for adjusting selectivity
with mobile phase manipulation. T he selectivity of a peptide separation
is different for mobile phases containing TFA and formic acid. T he use of
columns that give good performance with either modifier create an option
for developing methods in addition to the desirable impact on sensitivity
in MS detection. Furthermore, because peak shape and resolution are not
dependent on the presence of TFA, the concentration of TFA can be adjusted
to fine tune the selectivity of a peptide separation.
More dramatic changes in selectivity can be induced by changing the pH
of the separation from low pH to pH 7 or even pH 10 (e.g., using XBridge
BEH130 C18 columns).
Column Selectivity
Ion-Exchange Chromatography
T he retention mechanism of peptide separations is similar on reversedphase HPLC columns in general. T he selectivity of the separation is
significantly affected by the base sorbent, pore size, alkyl ligand
density, and residual silanol endcapping. T hese small differences in
separation selectivity can be a part of developing a peptide separation. If it is difficult to ac hieve a desirable separation on a particular
R P-HPLC column, then the use of an alternative R P-HPLC column
chemistry may be tested. For example, different separation selectivities
will be observed using XBridge BEH300 C18 compared to Atlantis T3.
Mobile-Phase Selectivity
Peptides are c harged molecules that do not freely interact with the
hydrophobic surface of reversed-phase columns. T he mobile phase
is typically, therefore, acidified to protonate the peptide carboxyl
groups, increasing retention. Trifluoroacetic acid is often preferred for
this purpose because it also forms ion pairs with the basic groups of
peptides. T he combination of protonation and ion pairing leads to the
202
Size-Exclusion Chromatography
Size-exclusion chromatography is sometimes used for peptide separations.
T he peptides are separated based on their size in solution. T he resolution
seldom compares favorably to either reversed phase or ion exchange, but
it can provide an alternative for some samples. T he composition of the
separation buffer will require careful adjustment for each particular peptide
www.waters.com
sample. Elevated ionic strength and the addition of some organic solvent,
typically methanol or propanol, may be necessary for good peak shape. T he
size-exclusion packings, particularly those with smaller pore sizes, described
in the section on protein chromatography, are suitable for peptides.
BioSuite PA-A column is especially suitable for polar peptides and for
use with MS-compatible eluents. T he BioSuite PA-B column has 300
pores for the analysis of large and hydrophobic peptides. Both BioSuite
Peptide Analysis columns are specifically QC-tested with protein digests
to ensure batc h-to-batc h product consistency and column-to-column
performance reproducibility.
Many other Waters columns have been used for particular peptide mixtures.
It is often observed that most peptides will give reasonable chromatography
on many different columns. T here is, however, often a particular column
that, for some subtle and unpredictable reason, works better than any other
for a particular pair of peptides.
Isolation of Natural and Synthetic Peptides
Purification of large amounts of individual peptides is most often achieved
with reversed-phase chromatography using the same principles and ideas
described above. T he columns are often c hosen to provide economical
separations on a larger scale so they are packed with larger size particles.
T he Peptide Separation Technology columns described above are available
in larger particle sizes and larger dimension columns for this purpose. T he
minimal secondary interactions mean that these columns can be used for
peptides representing a wide range of sizes and chemical properties. T he
surface chemistry of these packings is consistent and scalable across the
range of particle sizes. As necessary, separations may be optimized on
small scale columns and then transferred to columns large enough for the
required sample.
For some complex mixtures, two step separations may give the highest yield
and purity. In general, the best first step is cation-exchange chromatography
followed by a reversed-phase step for final isolation and desalting.
Proteomics, Analysis of Protein Expression, and Biomarker Discovery
For analyses used in proteomics and biomarker discovery, the available
samples are extremely limited so the requirements of c hromatographic
separation must be coupled with extreme sensitivity. T he sample
c hromatographic tools and principles described above are implemented
on the smallest scale in the nanoACQUIT Y UPLC system with nanoACQUIT Y columns. T he application solutions are described below in the
section on MS and LC/MS consumables.
Other Waters columns also have proven suitable for peptide mapping.
T he BioSuite Peptide Analysis columns are two premier reversed-phase
column c hemistries specifically optimized for peptide mapping. T he
www.waters.com
203
0.70
0.36
0.60
0.32
Average
0.24
0.20
AU
AU
0.40
Basic
pI 10.3
15 Residues
1809 da
0.28
pI 6.4
17 Residues
1772 da
0.50
0.30
0.16
0.12
0.20
0.08
0.10
0.04
0.00
0.00
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
15
90 min
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90 min
60
65
70
75
80
85
90 min
0.70
Large
0.60
Hydrophobic
0.32
pI 4.7
~
~40 Residues
~
~4000 da
0.50
pI 7.3
16 Residues
1872 da
0.28
0.24
0.40
AU
AU
0.20
0.30
Peptide
0.16
0.12
0.20
0.08
Preservative
0.10
0.04
0.00
0.00
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90 min
15
20
25
30
35
40
45
50
55
PST columns are compatible with a broad range of peptides. A single column can be used to purify the wide range of sample types generated in a peptide
synthesis laboratory. This minimizes the need to switch for columns separating peptides of extreme sizes, isoelectric point, or hydrophobicity.
204
www.waters.com
4 residue
25 residue
20 residue
507.3
2447.2
1565.8
1.0e-1
8.0e-2
AU
A
B
C
6.0e-2
4.0e-2
2.0e-2
0.0
20
25
30
35
40
45
50
55
60
65
70
75
80 min
75
80 min
6.0e-2
5.0e-2
AU
4.0e-2
3.0e-2
2.0e-2
1.0e-2
0.0
20
25
30
35
www.waters.com
40
45
50
55
60
65
70
205
Long-term Stability
100
Peptide Separation
Technology
B
90
C
% Initial Retention
80
D
70
60
50
50
100
2.2e-1
2.0e-1
AU
1.8e-1
1.6e-1
Length
(mm)
50
100
150
250
50
100
150
250
Length
(mm)
50
100
150
250
1.4e-1
1.2e-1
1.0e-1
8.0e-2
30
32
34
36
38
40
42
44
46
48
50 min
1.5
1.3
AU
1.1
9.0e-1
7.0e-1
5.0e-1
3.0e-1
i.d.
(mm)
10
10
10
10
19
19
19
19
1.0e-1
2
5.25e-2
10
11
12 min
i.d.
(mm)
30
30
30
30
4.75e-2
4.25e-2
3.75e-2
AU
3.25e-2
2.75e-2
2.25e-2
1.75e-2
1.25e-2
7.5e-3
2.5e-3
4
10
11
12
13
14
15
16 17 min
Waters PST columns, available in different particle size and column configurations,
are well-suited for various lab-scale purification needs.
206
www.waters.com
Inner
Diameter
75 m
75 m
75 m
75 m
100 m
150 m
75 m
100 m
150 m
75 m
75 m
75 m
Length
Particle
Size
Part No.
100 mm
150 mm
200 mm
250 mm
100 mm
100 mm
100 mm
100 mm
100 mm
150 mm
200 mm
250 mm
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
186003542
186003543
186003544
186003545
186003546
186003550
186003810
186003811
186003812
186003813
186003814
186003815
Inner
Diameter
Length
Particle
Size
Part No.
75 m
75 m
75 m
100 m
100 m
100 m
150 m
150 m
150 m
300 m
300 m
300 m
75 m
75 m
75 m
100 m
100 m
100 m
150 m
150 m
150 m
300 m
300 m
300 m
300 m
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
186003588
186003589
186003590
186003591
186003592
186003593
186003594
186003595
186003596
186003597
186003598
186003599
186003632
186003633
186003634
186003635
186003636
186003637
186003638
186003639
186003640
186003641
186003642
186003643
186003682
*NOTE: Use of Waters 300 m Capillary Inlet Tubing (Part Number 430001471) and 300 m Capillary Outlet Tubing (Part Number
430001472) required for use with 300 m I.D. capillary columns. Waters Kit Part Number 700002757 contains both the 300 m
Capillary Inlet Tubing (Part Number 430001471) and the 300 m Capillary Outlet Tubing (Part Number 430001472).
Inner
Diameter
Length
Particle
Size
2.1 mm
2.1 mm
2.1 mm
2.1 mm
100 mm
100 mm
100 mm
100 mm
1.7 m
1.7 m
3.5 m
3.5 m
www.waters.com
Part No.
186004896
186004897
186004904
186004905
Description
Inner
Diameter
Length
Particle
Size
Part No.
2.1 mm
2.1 mm
2.1 mm
2.1 mm
2.1 mm
2.1 mm
1.0 mm
1.0 mm
1.0 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
186003554
186003555
186003556
186003685
186003686
186003687
186005592
186005593
186005594
Part No.
186004629
186003975
205000343
700002775
Inner
Diameter
Length
Particle
Size
Part No.
1 mm
1 mm
1 mm
2.1 mm
2.1 mm
2.1 mm
2.1 mm
4.6 mm
4.6 mm
4.6 mm
4.6 mm
1 mm
1 mm
1 mm
2.1 mm
2.1 mm
2.1 mm
2.1 mm
4.6 mm
4.6 mm
4.6 mm
4.6 mm
1 mm
1 mm
1 mm
2.1 mm
2.1 mm
2.1 mm
2.1 mm
4.6 mm
4.6 mm
4.6 mm
4.6 mm
1 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
50 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
186003560
186003561
186003562
186003563
186003564
186003565
186003566
186003567
186003568
186003569
186003570
186003571
186003572
186003573
186003574
186003575
186003576
186003577
186003578
186003579
186003580
186003581
186003604
186003605
186003606
186003607
186003608
186003609
186003610
186003611
186003612
186003613
186003614
186003615
207
Inner
Diameter
Length
1 mm
1 mm
2.1 mm
2.1 mm
2.1 mm
2.1 mm
4.6 mm
4.6 mm
4.6 mm
4.6 mm
10 mm
10 mm
10 mm
10 mm
19 mm
19 mm
19 mm
4.6 mm
4.6 mm
4.6 mm
4.6 mm
10 mm
10 mm
10 mm
10 mm
19 mm
19 mm
19 mm
30 mm
30 mm
30 mm
30 mm
10 mm
10 mm
10 mm
10 mm
19 mm
19 mm
19 mm
4.6 mm
4.6 mm
4.6 mm
4.6 mm
10 mm
10 mm
10 mm
10 mm
19 mm
19 mm
19 mm
30 mm
30 mm
30 mm
30 mm
100 mm
150 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
50 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
50 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
208
Particle Configuration
Size
Style
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
Column
OBD Column
OBD Column
OBD Column
Column
Column
Column
Column
Column
Column
Column
Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
Column
Column
Column
Column
OBD Column
OBD Column
OBD Column
Column
Column
Column
Column
Column
Column
Column
Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
Part No.
186003616
186003617
186003618
186003619
186003620
186003621
186003622
186003623
186003624
186003625
186003582
186003583
186003584
186003585
186003586
186003587
186003945
186003648
186003649
186003650
186003651
186003652
186003653
186003654
186003655
186003656
186003657
186003658
186003659
186003660
186003661
186003662
186003626
186003627
186003628
186003629
186003630
186003631
186003946
186003663
186003664
186003665
186003666
186003667
186003668
186003669
186003670
186003671
186003672
186003673
186003674
186003675
186003676
186003677
Description
Inner
Diameter
Length
Particle Configuration
Size
Style
10 mm
19 mm
10 mm
19 mm
10 mm
19 mm
10 mm
19 mm
10 mm
10 mm
10 mm
10 mm
10 mm
10 mm
10 mm
10 mm
5 m
5 m
10 m
10 m
5 m
5 m
10 m
10 m
Guard Column
Guard Column
Guard Column
Guard Column
Guard Column
Guard Column
Guard Column
Guard Column
Part No.
186004469
186004468
186004465
186004464
186004471
186004470
186004467
186004466
Length
moles of a
Single Peptide
Weight of a Single
Peptide (mg)
10 mm
10 mm
10 mm
10 mm
19 mm
19 mm
19 mm
19 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
0.25-5
0.25-5
0.25-5
0.25-5
1-18
1-18
1-18
1-18
0.5-10
0.5-10
0.5-10
0.5-10
2.0-36
2.0-36
2.0-36
2.0-36
4.5-9
4.5-9
4.5-9
4.5-9
16-32
16-32
16-32
16-32
Description
Qty.
10 x 10 mm Cartridge Holder
19 x 10 mm Cartridge Holder
Replacement o-ring 7.8 mm
Replacement o-ring 10 mm
Replacement o-ring 19 mm
Part No.
289000779
186000709
2/pkg
2/pkg
2/pkg
700001019
700001436
700001020
www.waters.com
Part No.
Qty/Box
176001835
186004713
186003685
600001043
1
1
3
176001836
186004713
186003607
600001043
1
1
3
186001829
186002499
186002481
186002483
WAT058942
186001831
186003522
725000417
725000418
1
1
50
50
25
1
2
1
1
www.waters.com
Literature
References
209
2.1 x 100 mm
Sample:
Detection:
Run Time:
70 min
Temperature:
40 C
Solvent A:
Solvent A:
Gradient:
Time(min)
Flow Rate
%A
%B
Initial
0.200
95.0
5.0
1.00
0.200
95.0
5.0
59.00
0.200
50.0
50.0
60.00
0.200
30.0
70.0
61.00
0.200
95.0
5.0
Curve
Columns:
210
10
15
20
25
30
35
40
45 mIn
www.waters.com
Length
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
Configuration
Style
Column
Column
Column
Column
Column
Column
Column
Column
Column
Atlantis T3
C18, 3 m
186003713
186003714
186003717
186003718
186003719
186003721
186003722
186003723
C18, 5 m
186003730
186003731
186003734
186003735
186003736
186003738
186003739
186003740
ACQUITY HSS
T3, 1.8 m
C18, 1.8 m
1 pack
3 pack
1 pack
3 pack
1 pack
3 pack
186003535 176001127 186003529 176001121 186004114 176001556
186003536 176001129 186003530 176001122 186004115 176001557
186003537 176001130 186003531 176001123 186004116 176001558
186003538 176001131 186003532 176001124 186004118 176001560
186003539 176001132 186003533 176001125 186004119 176001561
186003540 176001133 186003534 176001126 186004120 176001562
186004679 176001814 186004683 176001818 186004687 176001822
186004680 176001815 186004684 176001819 186004826 176001823
186004681 176001816 186004685 176001820 186004689 176001824
Inner
Diameter
Length
Configuration
Style
Particle
Size
Part No.
10 mm
10 mm
10 mm
10 mm
10 mm
19 mm
19 mm
19 mm
19 mm
19 mm
30 mm
30 mm
30 mm
30 mm
50 mm
50 mm
50 mm
10 mm
10 mm
10 mm
19 mm
19 mm
19 mm
19 mm
30 mm
30 mm
50 mm
50 mm
50 mm
50 mm
10 mm
50 mm
100 mm
150 mm
250 mm
10 mm
50 mm
100 mm
150 mm
250 mm
50 mm
75 mm
100 mm
150 mm
50 mm
100 mm
150 mm
10 mm
150 mm
250 mm
10 mm
50 mm
150 mm
250 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
Guard
Column
Column
Column
Column
Guard
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
Guard
Column
Column
Guard
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
OBD Column
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
10 m
1860036951
186003691
186003692
186003693
186003694
1860036992
186003696
186003697
186003698
186004026
186003700
186003701
186003702
186003703
186004080
186004081
186004082
1860037061
186003704
186003705
1860037102
186003707
186003708
186003709
186003711
186003712
186004083
186004084
186004085
186004086
ACQUITY HSS
ACQUITY HSS
Description
Qty.
10 x 10 mm Cartridge Holder
19 x 10 mm Cartridge Holder
Replacement o-ring 7.8 mm
Replacement o-ring 10 mm
Replacement o-ring 19 mm
Part No.
289000779
186000709
2/pk
2/pk
2/pk
700001019
700001436
700001020
www.waters.com
211
Eluent A:
Eluent B:
Flow rate:
Gradient:
Injection Volume:
Temperature:
Detection:
%B
0
28
Batch 112
Batch 113
Batch 114
Ideal choice for LC/MS applications using formic acid (FA) that
minimizes ion-suppression
212
10
15
20
25
30
35
40
45
50
55 Min.
Inner
Diameter
Length
BioSuite C18
3 m PA-A
BioSuite C18
3.5 m PA-B
2.1 mm
2.1 mm
2.1 mm
2.1 mm
4.6 mm
4.6 mm
4.6 mm
4.6 mm
50 mm
100 mm
150 mm
250 mm
50 mm
100 mm
150 mm
250 mm
186002425
186002426
186002427
186002428
186002429
186002430
186002431
186002432
186002433
186002434
186002435
186002436
186002437
186002438
186002439
186002440
www.waters.com
Column:
Eluent:
Flow rate:
Temperature:
Detection:
III
II
[Asn1, Val5]-II
Des-Asp1, [IIe8]-II
Exclusion Limit
(Daltons) against
Polyethylene
Inner
Glycol
Diameter
Pore
Size
Description
Matrix
Polymer 1300
Polymer
n/a
Polymer 1000
Polymer 1000
>4,000,000
500
1,000,000
1,000,000
4.6 mm
4.6 mm
7.5 mm
21.5 mm
10
15
20
25 Min.
Length
Column
Volume
(mL)
# Approx Protein
Binding Capacity
Per Pre-Packed
Column
Part No.
50 mm
35 mm
75 mm
150 mm
0.83
0.58
3.31
54.45
58 mg*
2.9 mg**
132 mg**
2,178 mg**
186002182
186002183
186002184
186002185
pore size materials (100 and 300) with a C18 or C4 bonded phase.
For more information and part numbers, visit waters.com/biosep.
Physical Characteristics
Packing
Delta-Pak
Chemistry
Particle
Size
Particle
Shape
Pore
Size
Carbon
Load
Endcapped
C18
C18
5 m
5 m
Spherical
Spherical
100
300
17%
7%
Yes
Yes
C4
C4
5 m
5 m
Spherical
Spherical
100
300
7%
3%
Yes
Yes
C18
C18
15 m
15 m
Spherical
Spherical
100
300
17%
7%
Yes
Yes
C4
C4
15 m
15 m
Spherical
Spherical
100
300
7%
3%
Yes
Yes
Column:
Part Number:
Sample:
Injection:
Mobile Phase:
Gradient:
Conditions:
Flow Rate:
Detection:
Description
Waters Insulin HMWP Column
Protein-Pak 125 Sentry Guard Column,
2/pkg (requires holder)
Sentry Universal Guard Column Holder
Dimensions
Part No.
7.8 x 300 mm
WAT201549
3.9 x 20 mm
186000926
WAT046910
Tested to perform in the method published in PharmaEuropa Vol. 8, No 3, pages 359-360, September 1996.
www.waters.com
213
Sample:
Mobile Phase:
13
56
23
12
8
11
In the past youve had no choice but to deal with variability by finding
costly and time-consuming ways around this problem. The regulatory guidelines being adopted worldwide as a result of the International Conference
on Harmonization (ICH) and the U.S. Food and Drug Administrations
specified biological products initiatives are placing increasingly stringent
demands on this process. Full analytical characterization is becoming
more important for regulatory filing, making column variability an
unacceptable risk.
Gradient:
Injection:
Flow Rate:
Temperature:
Detection:
10
As a scientist, you know that tremendous time and resources are required
when developing HPLC assays for well-characterized biopharmaceuticals.
You also know that column-to-column variability is the Achilles heel of
this demanding process.
A. Symmetry C18, 5 m,
100, 4.6 x 150 mm
B. Symmetry300 C18, 5 m,
300, 4.6 x 150 mm
Tryptic digests of cytochrome c (bovine)
Solvent A: 0.1% TFA in water
Solvent B: 0.1% TFA in acetonitrile
10
13
5,6
B
2
3
4
7
12
20
11
40 Min.
Length
Particle
Size
C18
C4
1.0 mm
2.1 mm
2.1 mm
2.1 mm
4.6 mm
4.6 mm
4.6 mm
4.6 mm
2.1 mm
3.9 mm
4.6 mm
4.6 mm
4.6 mm
19 mm
19 mm
19 mm
150 mm
50 mm
100 mm
150 mm
50 mm
75 mm
100 mm
150 mm
150 mm
150 mm
50 mm
150 mm
250 mm
10 mm
50 mm
100 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
186000185
186000187
186000188
186000200
186000201
186000189
186000190
186000197
WAT106172
WAT106154
WAT106209
WAT106157
WAT106151
186001847
186001848
186001849
186000276
186000277
186000278
186000279
186000280
186000281
186000282
186000283
186000285
186000286
186000287
186000288
186000289
214
www.waters.com
Hydrophobic-Interaction Chromatography
Ion-Exchange Chromatography
In ion-exc hange separations, the distribution and net c harge on the
proteins surface determines the interaction of the protein with the
c harged groups on the packing materials surface. T he c harges on the
protein and the packing material must be opposite for interaction to
occur. T he variables that can be manipulated, suc h as buffer pH, buffer
composition, gradient slope, and gradient forming salt, offer a wide
range of options for optimizing a separation.
Size-Exclusion Chromatography
T he apparent size and shape of proteins in solution is the basis for
separation in size-exclusion c hromatography. T his simple separation mec hanism relies on protein size differences, packing material,
pore size, and particle size as well as the operating conditions, suc h
as sample concentration and volume for the successful resolution of
protein mixtures. Size-exclusion packing materials have evolved from
the large particle polysaccharide soft gels to small, rigid packings that
offer enhanced resolution and significantly faster separations.
Size exclusion has a wide range of applicability in protein isolations. It is
often used to assay fractions, as run times are relatively short, and the
technique relatively straightforward. It can also be used for buffer exchange,
as the low molecular weight salt molecules are easily separated from the
much larger proteins.
Reversed-Phase Chromatography
Reversed phase is an alternative hydrophobic-separation technique that
relies on the interaction between the proteins non-polar amino acid side
chains with the packing surface and ligands. T his mechanism provides
another high resolution dimension for protein chromatography that can
yield good recovery of biological activity with lower molecular weight
proteins and peptides. It is an excellent tool for assessing fraction purity
and for preparing proteins for sequencing or structural analysis.
Affinity Chromatography
Affinity is the only chromatographic mode in which proteins are isolated
on the basis of their biospecific interaction with an immobilized ligand.
Proteins can be recovered with retention of bioactivity and high purity.
Modern affinity packings take advantage of rigid base materials that
combine enhanced bed stability for crude sample processing with high
throughput for rapid microgram to gram scale affinity purifications. Many
separations can be achieved in a single step resulting in more costeffective separations.
www.waters.com
Sample Preparation
Protein samples derived from tissue homogenates, cell lysis, or physiologic
fluids frequently require removal of particulate contaminants prior to LC,
MS, or LC/MS analysis. In some situations, use of an on-line protein desalting device (e.g., MassPREP on-line desalting cartridge) can effectively be
used to remove ion-suppressing compounds prior to mass determination by
LC/MS techniques. Other situations exist where simple filtration or centrifugation can be effectively used to remove insoluble components prior to
analysis. Glycoproteins offer additional challenges that can be addressed
via the use of specialty reagents such as Waters RapiGest SF surfactant
that has been shown to significantly enhance protein deglycosylation. In
addition, solid-phase extraction techniques, such as those offered in Waters
MassPREP Deglycosylation Kit and Plate, can effectively isolate and concentrate glycans, cleaved from gylcoproteins, prior to analysis. Accell Plus
ion-exc hanger cartridges are also available for solid-phase extraction of
proteins from complex matrices or for isolation or sample concentration.
215
Hydrophobic Interaction
Affinity
Reversed Phase
Size Exclusion
Concentrates sample
Concentrates sample
Concentrates sample
Concentrates sample
Rapid
Superb selectivity
Moderate resolution
High capacity
High capacity
Widespread applicability
Outstanding resolution
High capacity
Good recovery of activity
Simple technique
Moderate capacity
Significant purification
in a single step
Hydrophobic Interaction
Affinity
Reversed Phase
Size Exclusion
Ligand leakage
Moderate resolution
Non concentrating
Moderate capacity
For instance, many of the initial feed streams for protein isolations are
relatively dilute protein solutions. Chromatographic modes that concentrate
samples (ion exchange, hydrophobic interaction, and affinity) are good starting points for these solutions.
For samples such as cell lysates and homogenates or physiologic fluids,
there are additional considerations. T hese mixtures contain cell fragments,
lipids, and nucleic acid fragments, which will bind strongly to a column.
Columns becoming contaminated in this way are typically cleaned using
0.1 M -1.0 M NaOH prior to re-equilibration with the starting buffers.
T his would destroy a peptide or protein ligand on an affinity column
and would irreversibly damage a silica-based, reversed-phase column.
T herefore, ion exchange or hydrophobic interaction would be appropriate
modes for a first purification step.
In reversed-phase chromatography, the sample is loaded onto the column
in an aqueous environment and then eluted using an increasing gradient of
an organic solvent, such as acetonitrile or propanol. T he major drawback to
this mode of chromatography is that some proteins will denature (causing
loss of biological activity) in the presence of the organic solvents at the
concentration required to elute the protein. As a general rule, the likelihood
of denaturation increases with increasing molecular weight of the protein.
216
www.waters.com
BEH Technology
E tO C H 2 C H 2 O E t
Si O
Si
O
O
Si
E tO
OE t
Si O
Si
Et
O
Si
O Et
O Et
OEt
E tO
E tO
Si
E tO
O Et
E tO
E tO S i
E tO
Polyethoxysilane
(BPEOS)
Tetraethoxysilane
(TEOS)
Bis(triethoxysilyl)ethane
(BTEE)
www.waters.com
O Et
CH 2
Si O Et
CH 2
O Et
217
Based upon the highly successful BEH particle technology, the new patent
pending ACQUIT Y UPLC BEH200 SEC column represents the first
commercially available sub 2 m SEC chemistry. Fully optimized for the
analysis of proteins and their aggregates with molecular weights ranging
mAb monomer
mAb monomer
Aggregates
10.3%
Aggregates
10.7%
Column:
Sample:
HPLC system:
Eluent:
Flow Rate:
Injection:
Wavelength:
10
15
20
25
30
35
40 Min.
Column:
6 Min.
Sample:
UPLC system:
Eluent:
Flow Rate:
Injection:
Wavelength:
10000000
1000000
MW
100000
Ribonuclease A
(~13,700 Da)
10000
1000
100
10
1
0
0.5
1.5
2.5
Elution Volume
218
www.waters.com
Effect of Eluent Ionic Strength on the SEC Analysis of the Basic Protein Lysozyme on 100% Silica vs BEH SEC Particles
Flow rate:
Mobile phase:
0.3 mL/min
10, 25 or 100 mM
sodium phosphate, pH 6.8
Lysozyme, 10 mM
10
14
18
22
26
30
34
38
42
50 Min.
46
10
11
12
13
15 Min.
14
T he combination of the BEH base particle and the patent pending innovative
diol bonding process, results in column stability, performance and lifetime
unheard of in traditional size exclusion chromatographic columns.
ACQUITY UPLC BEH200 SEC Particle and DIOL Bonding Technology Provides a Stable Chemistry with Outstanding Column Life
Sample:
Flow rate:
Mobile phase:
Lysozyme
0.4 mL/min
25 mM sodium
phosphate, pH 6.8
with 0.15 m sodium chloride
11
12
13
14
15 Min.
www.waters.com
10 Min.
219
Batch A
Analyte
pI
MW
4.6
669,000
2. IgG, 2 mg/mL
6.7
150,000
4.6
66,400
6.8, 7.2
17,000
N/A
112
1
5
10 Min.
Batch B
2
3
1
5
10 Min.
Batch C
2
3
Description
ACQUITY UPLC BEH200, 1.7 m SEC
ACQUITY UPLC BEH200, 1.7 m SEC
Dimensions
Part No.
4.6 x 150 mm
4.6 x 300 mm
186005225
186005226
10 Min.
Literature
References
Protein Separation Technology Brochure,
Literature Reference 720002577EN
Protein-Pak Hi Res Ion Exchange
(IEX) Columns for ACQUITY UPLC
Applications Solution Brochure,
Literature Reference 720003346EN
ACQUITY UPLC SEC System Solution Brochure,
Literature Reference 720003398EN
220
www.waters.com
Tolerates extreme pH
and temperature
Batch-to-Batch Reproducibility
Sample:
MassPREP Protein Std (Part No. 186004900)
0.1% TFA in 5% acetonitrile
1. Ribonuclease A (0.04 mg/mL)
2. Cytochrome c (0.06 mg/mL)
3. Bovine Serum Albumin (0.20 mg/mL)
4. Myoglobin (0.13 mg/mL)
5. Enolase (0.22 mg/mL)
6. Phosphorylase b (0.59 mg/mL)
Column:
Eluent A:
Eluent B:
Flow Rate:
Gradient:
Column Temp.:
Detection :
Sample:
MassPREP Protein Std (Part No. 186004900)
0.1% TFA in 5% acetonitrile
1. Ribonuclease
2. Cytochrome c
3. BSA
4. -Lactoglobulin
5. Enolase
6. Phosphorylase b
5
4
0.15
Batch A
0.10
AU
3
6
Batch B
0.05
Batch C
0.00
12
18 min
BEH300 C4 columns can be used with proteins that have a wide range of properties. This protein mix was
chosen to represent a range of isoelectric points, molecular weights, and hydrophobicities.
www.waters.com
10
12
14
16
18
20
22 Min.
This comparison shows the consistent batch-to-batch performance with a protein separation.
221
Ovalbumin
0.05
Sample:
Columns:
AU
Eluent A:
Eluent B:
Flow Rate:
Gradient:
0.00
0
10
20
30
40
50
60
70
80
90
100
110
120
Column Temp.:
Detection :
130 min
Column carryover was tested by running multiple gradients following a single injection. Protein peaks observed during the first gradient are not found in subsequent gradients.
Mobile Phase A:
Mobile Phase B:
Flow Rate:
Gradient:
Column Temp.:
Injection:
Detection :
AU
0.24
4
2
Column:
Eluent A:
Eluent B:
Flow Rate:
Gradient:
Fd
LC
Fc/2
6
%
0.12
Sample:
Column Temp.:
BEH300 C4
3.5 m
0.36
0.00
AU
0.36
BEH300 C4
1.7 m
0.24
Fc/2
Clip 2
1
0.12
0.00
Fc/2
Clip 1
12
24
36
48
60 min
The protein test mixture was separated on two BEH300 C4 columns, one with 3.5 m particles and the other with
1.7 m particles. The UPLC separation provides sharper peaks for all the proteins in the test mixture. This translates
into better resolution as shown by the multiple peaks around phosphorylase, at approximately 50 min. This comparison, with both columns, was performed on a UPLC system to preserve the minimized band-broadening. The benefits of
the small particle UPLC BEH300 C4 column would be lost without the optimized ACQUITY UPLC System.
10
12
14
16
18
20
22
24
26
28
30
32
34 Min
T he large fragments obtained through LysC digestion of a monoclonal antibody can be separated on the UPLC
BEH300 C4 column coupled directly to ESI/Tof MS for identification of the individual peptide products.
Waters BEH300 C4 Offerings for Traditional HPLC and Advanced UPLC Protein Separations
UPLC Columns
Particle Size
Dimensions
Part No.
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
1 x 50 mm
1 x 100 mm
1 x 150 mm
186004495
186004496
186004497
186005589
186005590
186005591
1.7 m
VanGuard Pre-Column
186004623
Note: ACQUIT Y UPLC BEH300 C4, 1.7 m columns are designed for
use with the ACQUITY UPLC system. T he benefits of the small particle
packing in ACQUITY UPLC BEH300 C4, 1.7 m columns are only realized
with the low system volume and low detector dispersion of an ACQUITY
UPLC system.
222
HPLC Columns
XBridge BEH300 C4
XBridge BEH300 C4
XBridge BEH300 C4
XBridge BEH300 C4
XBridge BEH300 C4
XBridge BEH300 C4
XBridge BEH300 C4
XBridge BEH300 C4
Custom BEH300 C4
ACQUITY BEH300 C4
Method Validation Kit*
ACQUITY UPLC BEH300 C4
ACQUITY UPLC BEH300 C4
ACQUITY UPLC BEH300 C4
Particle Size
Dimensions
Part No.
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
2.1 x 50 mm
2.1 x 100 mm
2.1 x 150 mm
2.1 x 250 mm
4.6 x 50 mm
4.6 x 100 mm
4.6 x 150 mm
4.6 x 250 mm
186004498
186004499
186004500
186004501
186004502
186004503
186004504
186004505
186004506
1.7 m
2.1 x 100 mm
186004899
1.7 m
1.7 m
1.7 m
1.0 x 50 mm
1.0 x 100 mm
1.0 x 150 mm
186005589
186005590
186005591
www.waters.com
columns
batch consistency.
Column:
Flow Rate:
Eluent A:
Eluent B:
Gradient:
Temperature:
Detection:
0.040
0.030
0.020
0.010
0.000
0.040
0.030
Column:
Flow Rate:
Eluent A:
Eluent B:
Gradient:
0.020
Humanized IgG1
0.010
0.000
0.020
0.008
Humanized IgG2
0.010
0.006
0.000
0.004
0.015
0.002
0.000
K1
K0
0.010
K2
Chimeric IgG1
0.005
10
12
14
16
18
20
22
24
26
28
30 min
0.000
Cation-Exchange chromatography is a useful tool for the characterization and quantitation of mAb or
recombinant protein variants. Use of Waters Protein-Pak Hi Res CM column on an ACQUITY UPLC System
increases sample throughput while maintaining resolution between intended product and undesired variants.
www.waters.com
10
12
14
16
18
20
22
24 min
Sequence, production, storage, and shipping conditions influence the degree of variants contained in a biotherapeutic
protein. Waters Protein-Pak Hi Res CM column can successfully resolve variations that may involve as little as a
single amino acid change (K0 = No terminal lysines, K1 = One terminal Lysine, and K2 = Two terminal Lysines).
223
Protein-Pak Hi Res Q, 5 m,
4.6 x 100 mm
0.6 mL/min
20 mM Tris Buffer, pH 8.5
20 mM Tris Buffer, pH 8.5, 0.5 M NaCl
0-60% B in 30 min
30 C
280 nm
Flow Rate:
Mobile Phase A:
Mobile Phase B:
Gradient:
Temperature:
Detection:
0.02
0.01
Batch A
0.00
0.02
0.01
Myoglobin
Apo-Transferrin
Batch B
Description
Protein-Pak
Hi Res Q
Protein-Pak
Hi Res CM
Protein-Pak
Hi Res SP
Ion Exchange
Strong Anion
Weak Cation
Strong Cation
Quaternary
ammonium
Carboxymethyl
Sulfopropyl
Hydrophilic polymer
Hydrophilic polymer
Hydrophilic polymer
Pore size
Non porous
Non porous
Non porous
i.d. x L (mm)
4.6 x 100
4.6 x 100
4.6 x 100
Counter ion
Cl-
Na +
Na +
Functional Group
Matrix
Particle size (m)
pH range
Temperature (C)
pKa
Flow Rates
3-10
3-10
3-10
10-60
10-60
10-60
10.5
4.9
2.3
0.3-0.6 mL/min
0.5-1.4 mL/min
0.5-1.4 mL/min
58
33
25
0.02
Trypsin Inhibitor
0.01
Batch C
0.00
3
10
11
12
13 min
Each batch of Protein-Pak Hi Res SP, CM, and Q column packing material is chromatography tested using a relevant
protein standard mixture to help ensureconsistent and predictable performance (Protein-Pak Hi Res Q data shown).
Description
Protein-Pak Hi Res CM, 7 m
Protein-Pak Hi Res SP, 7 m
Protein-Pak Hi Res Q, 5 m
Dimensions
Part No.
4.6 x 100 mm
4.6 x 100 mm
4.6 x 100 mm
186004929
186004930
186004931
Note: Only when Protein-Pak Hi Res IEX columns are combined with the
ACQUIT Y UPLC system are the full performance benefits realized. See
Waters service notes P/N 715002147A for ACQUIT Y UPLC system
configuration guidelines for ion-exchange chromatography.
More information regarding this and additional columns for protein sample
preparation and analysis can be found at www.waters.com/Proteins.
For optimal resolution of complex samples, do not exceed 20% of the columns protein binding capacity.
224
www.waters.com
Protein Selectivity Differences on BioSuite CM (Weak CationExchange) vs. SP (Strong Cation-Exchange) Columns
A) BioSuite CM,
10 m CXC
A) BioSuite SP,
2.5 m NP CXC
2
4
B) BioSuite SP,
10 m CXC
1
B) BioSuite SP,
10 m CXC
3
1
0
Sample:
Columns:
Eluent: A:
Eluent B:
Flow Rate:
Gradient:
Temperature:
Detection:
10
5
2
20
30
40 Min.
Description
BioSuite Q-PEEK 10 m AXC
BioSuite SP-PEEK 7 m CXC
4 3
Sample:
Columns:
Eluent: A:
Eluent B:
Flow Rate:
Gradient:
Temperature:
Detection:
Matrix
Pore
Size
Polymer 4000
Polymer 1300
5 4
6 Min.5
6 20
Min.
20 30
30
40 Min.
40 Min.
Exclusion Limit
(Daltons) against
Polyethylene
Inner
Glycol
Diameter
Length
# Approx Protein
Column Binding Capacity
Volume Per Pre-Packed
(mL)
Column
Part No.
>5,000,000
>4,000,000
4.6 mm
4.6 mm
50 mm
50 mm
0.83
0.83
58 mg*
58 mg**
186002176
186002182
n/a
n/a
500
500
4.6 mm
4.6 mm
35 mm
35 mm
0.58
0.58
2.9 mg*
2.9 mg***
186002179
186002183
BioSuite Q 10 m AXC
BioSuite Q 13 m AXC
BioSuite DEAE 10 m AXC
BioSuite DEAE 13 m AXC
Polymer
Polymer
Polymer
Polymer
1000
1000
1000
1000
1,000,000
1,000,000
1,000,000
1,000,000
7.5 mm
21.5 mm
7.5 mm
21.5 mm
75 mm
150 mm
75 mm
150 mm
3.31
54.45
3.31
54.45
331 mg*
5,445 mg*
99 mg*
1633 mg*
186002177
186002178
186002180
186002181
BioSuite SP 10 m CXC
BioSuite SP 13 m CXC
BioSuite CM 10 m CXC
BioSuite CM 13 m CXC
Polymer
Polymer
Polymer
Polymer
1000
1000
1000
1000
1,000,000
1,000,000
1,000,000
1,000,000
7.5 mm
21.5 mm
7.5 mm
21.5 mm
75 mm
150 mm
75 mm
150 mm
3.31
54.45
3.31
54.45
132 mg***
2,178 mg***
149 mg***
2,450 mg***
186002184
186002185
186002186
186002187
* Data generated with BSA ** Data generated with Gamma Globulin *** Data generated with Hemoglobin
# Note: For best resolution of complex samples, do not exceed 20% of the columns protein binding capacity
226
www.waters.com
Particle
Type
Pore
Size
Size
Dimensions
Part No.
Protein-Pak
Q 8HR
Polymeric strong
anion exchanger
8 m
1000
5 x 50 mm
5 x 100 mm
10 x 100 mm
5 x 50 mm
10 x 100 mm
5 x 50 mm
5 x 100 mm
10 x 100 mm
5 x 50 mm
5 x 100 mm
10 x 100 mm
5 x 50 mm
5 x 100 mm
10 x 100 mm
10 x 100 mm
WAT039575
WAT039630
WAT035980
WAT039782
WAT037663
WAT039791
WAT039783
WAT035650
WAT039780
WAT039786
WAT038564
WAT039570
WAT039625
WAT035655
WAT038567
5 x 50 mm
5 x 100 mm
10 x 100 mm
5 x 50 mm
WAT039790
WAT039785
WAT035970
WAT039787
Protein-Pak
Q 15HR
Protein-Pak
DEAE 8HR
Polymeric strong
anion exchanger
Polymeric weak
anion exchanger
15 m
1000
8 m
1000
Protein-Pak
DEAE 15HR
Polymeric weak
anion exchanger
15 m
1000
Protein-Pak
SP 8HR
Polymeric strong
cation exchanger
8 m
1000
Protein-Pak
SP 15HR
Protein-Pak
CM 8HR
Polymeric strong
cation exchanger
Polymeric weak
cation exchanger
15 m
1000
8 m
1000
Protein-Pak
CM 15HR
Polymeric weak
cation exchanger
15 m
1000
2
3
5
4
40 Min.
Q**
60 mg
130 mg
500 mg
Protein-Pak HR Packing
DEAE**
SP***
40 mg
150 mg
300 mg
40 mg
80 mg
300 mg
CM***
Dimensions
Part No.
7.5 x 75 mm
8 x 75 mm
21.5 x 150 mm
7.5 x 75 mm
8 x 75 mm
WAT088044
WAT011783
WAT010640
WAT088043
WAT011784
25 mg
45 mg
180 mg
Protein-Pak Q HR **
Type of material
Polymer
Protein binding capacity
60 mg/mL
Ion-exchange capacity
200 eq/mL
Nominal pK
11.7
Typical protein recovery
>95%
Typical recovery of biological activity >90%
pH stability
2-12
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Protein-Pak DEAE HR **
Protein-Pak CM HR ***
SP HR ****
Polymer
40 mg/mL
250 eq/mL
9.0
>95%
>90%
2-12
Polymer
25 mg/mL
175 eq/mL
5.7
>95%
>90%
2-12
Polymer
40 mg/mL
225 eq/mL
2.2
>95%
>90%
2-12
**
***
****
227
Dimensions
Bed Volume
(mL)
Flow Rate
(mL/min)
Pressure Rating
(psi/MPa)
Part No.
5 x 50 mm
5 x 100 mm
10 x 100 mm
10 x 200 mm
10 x 300 mm
10 x 600 mm
20 x 100 mm
20 x 200 mm
20 x 300 mm
20 x 600 mm
50 x 100 mm
50 x 200 mm
50 x 300 mm
50 x 600 mm
0.8-1.2
1.8-2.2
5-8
13-16
21-24
45-48
22-31
53-62
85-94
179-188
137-196
333-392
530-589
1118-1177
0-4
0-4
0-4
0-4
0-4
0-4
4-16
4-16
4-16
4-16
16-100
16-100
16-100
16-100
WAT064-01
WAT064-02
WAT021901
WAT021902
WAT021903
WAT021906
WAT027501
WAT027502
WAT027503
WAT027506
WAT023331
WAT023332
WAT023333
WAT023336
Part No.
Glass Tube
5 x 50 mm
5 x 100 mm
5 x 50 mm
5 x 100 mm
WAT038802
WAT038803
WAT038804
WAT038805
WAT038806
Filters, 10/pk
O-rings, 13/pk
(includes 10 inlet/outlet and 3 funnel)
Inlet Connector Assembly
Description
Dimensions
Part No.
Glass Tube
20 x 100 mm
20 x 200 mm
20 x 300 mm
20 x 100 mm
20 x 200 mm
20 x 300 mm
WAT019891
WAT019892
WAT019893
WAT027542
WAT027543
WAT027544
WAT027528
WAT027530
Plastic Shield
Description
Column Jacket
WAT038807
O-rings, 5/pk
Filters, 2/pk
Replacement Tubing (Tefzel)
1/8 inch o.d. x 0.040 inch i.d. x 10 feet
(3.2 mm o.d. x 1.02 mm i.d. x 3 m)
Inlet Connector Assembly
Distributors/inserts, 5/pk
WAT023344
WAT027525
700004715
WAT038800
Description
Dimensions
Part No.
Description
Dimensions
Part No.
Glass tube
10 x 100 mm
10 x 200 mm
10 x 300 mm
10 x 600 mm
10 x 100 mm
10 x 200 mm
10 x 300 mm
10 x 600 mm
WAT021992
WAT022033
WAT022034
WAT022035
WAT021927
WAT021945
WAT021946
WAT021947
WAT021907
WAT021910
Glass Tube
50 x 100 mm
50 x 200 mm
50 x 300 mm
50 x 100 mm
50 x 200 mm
50 x 300 mm
50 x 600 mm
WAT019876
WAT019877
WAT019878
WAT023370
WAT023371
WAT023372
WAT023373
WAT023345
WAT023343
WAT023344
Plastic shield
O-rings, 5/pk
Filters, 10/pk
Replacement tubing (Tefzel)
1/16 inch o.d. x 0.009 inch i.d. x 10 feet
(1.6 mm o.d. x 0.23 mm i.d. x 3 m)
Inlet Connector Assembly
WAT021950
Plastic Shield
O-rings, 5/pk
Filter, 2/pk
Replacement Tubing (Tefzel)
1/8 inch o.d. x 0.040 inch i.d. x 10 feet)
(3.2 mm o.d. x 1.02 mm i.d. x 3 m)
Inlet Connector Assembly
WAT023349
WAT021904
Consult your appropriate Care and Use Manual for additional spare parts information at www.waters.com/chemcu
228
www.waters.com
Description
1 Collet and nut assembly (3/8-24)
2 Ferrule 1/8 tube
3 Union 3/8-24 x Z fitting
Qty.
Part No.
10/pk
10/pk
5/pk
WAT005138
WAT005136
WAT005137
1/pk
1/pk
WAT082708
WAT082745
Qty.
Part No.
10/pk
10/pk
1/pk
WAT005138
WAT005136
WAT082734
Description
Qty.
Part No.
1/pk
1/pk
WAT082708
WAT021951
Description
Collet and nut assembly (3/8-24)
Ferrule 1/8 tube
Union 3/8-24 x Z fitting
Compression screw and ferrule Z fitting, plastic
Compression screw Z fitting, steel
Ferrule 1/16 steel
www.waters.com
Part No.
Description
1
2
3
4
5
6
Qty.
Description
Qty.
Part No.
10/pk
10/pk
5/pk
1/pk
10/pk
10/pk
WAT005138
WAT005136
WAT005137
WAT082708
WAT005070
WAT005063
Description
1 Collet and nut assembly (3/8-24)
2 Ferrule 1/8 tube
Qty.
Part No.
10/pk
10/pk
WAT005138
WAT005136
229
Particle
Size
Accell Plus CM *
40 m
PrepPak 1000 Module
Pore
Size
Part No.
300
WAT036545
WAT089592
Condition/
Equilibrate
Load Sample
(Black)
3
Step
Elute 1
Step
Elute 2
Step
Elute 3
For best resolution do not exceed 20% of the protein binding capacity.
**
Bovine serum albumin in 20 mM Tris/Cl pH 7.0 was used to measure protein binding capacity
of Accell Plus QMA.
***
Particle
Description
Particle
Size
Pore
Size
40 m
300
40 m
300
230
Qty.
Part No.
100 g
500 g
100 g
500 g
WAT010742
WAT010741
WAT010740
WAT010739
Sep-Pak Plus cartridges packed with Accell Plus ion exchangers provide a
rapid, economical means to clean up heavily contaminated samples that
would damage a high resolution column. They can also be used to rapidly
screen chromatographic conditions. These are also available in a variety
of configurations.
Description
Accell Plus CM
Accell Plus QMA
Ion-Exchange Type
Part No.
WAT020550
WAT020545
www.waters.com
BioSuite Ultra High Resolution (UHR), High Resolution (HR), and Standard
size-Exclusion Column packings use a rigid yet wettable silica-based
media that is stable from pH 2.5-7.5. As indicated in the calibration curve
tables, the exclusion limits of BioSuite SEC packings are determined by
the particle and pore size of the silica-based material. Particle size of the
SEC packing media as well as column length are important parameters that
100
TIC
66420
%
66443
%
66548
62801
68030
mass
60000
80000
100000
120000
140000
132835
100
0
10
12
14 Min.
132991
LC/MS:
Sample:
Column:
Injection:
Mobile Phase:
Flow Rate:
Temp:
MS Source:
MS Capillary (kV):
MS Cone (V):
Mass Range:
66417*
%
131011
66506
66555
65504
64540
60000
133095
133175
135141
67505
67648
70000
80000
90000
100000
110000
120000
130000
mass
140000
LC/MS Analysis of Protein Standards Using BioSuite 250, 5 m High Resolution (HR) SEC Column (LC/MS conditions as above)
4
100
6
3
1.
2.
3.
4.
5.
6.
T hyrogobulin
BSA Dimer
BSA
-Lactoglobulin
Myoglobin
Cytochrome C
669 KDa
136 KDa
66 KDa
36 KDa
16 KDa
12 KDa
2
1
10
12
14
16
18
20 Min.
BioSuite SEC Reference: SEC- MS Analysis of Aggregates in Protein Mixtures. Application Book Supplement
of LC/GC Europe. Sept. 2003. (Waters Literature Code Number: 720000743EN)
www.waters.com
231
Sample:
BioSuite 250,
4 m UHR SEC
Columns:
Eluent:
Flow rate:
Temperature:
Detection:
Molecular
MWWeight
100,000
10,000
BioSuite 125,
4 m UHR SEC
Column
Globular Protein
Mol. Wt. Range
Branched
Dextrans
Linear
PEG/PEO
BioSuite 125
BioSuite 250
5,000-150,000
10,000-500,000
1,000-30,000
2,000-70,000
500-15,000
1,000-35,000
1,000
Note: Operating flow rates for BioSuite Ultra-High Resolution (UHR) SEC columns (4.6 mm i.d.) are from
0.1 - 0.4 mL/min. Use of an HPLC system (e.g. Waters Alliance HPLC System) capable of operating at these
flows is essential for optimal UHR SEC column performance.
100
6
10
12 MIn.
Protein Calibration Curves for BioSuite High Resolution (HR) SEC Columns
Sample:
BioSuite 250,
5 m HR SEC
10 6
BioSuite 450,
8 m HR SEC
Eluent:
Flow rate:
Temperature:
Detection:
10 5
Molecular Weight
Columns:
10 4
Column
BioSuite 125,
5 m HR SEC
BioSuite 125
BioSuite 250
BioSuite 450
10 3
Globular Protein
Mol. Wt. Range
Branched
Dextrans
Linear
PEG/PEO
5,000-150,000
10,000-500,000
20,000-7,000,000
1,000-30,000
2,000-70,000
4,000-500,000
500-15,000
1,000-35,000
2,000-250,000
10 2
232
10
12 MIn.
www.waters.com
Sample:
10 6
Columns:
Eluent:
Flow rate:
Temperature:
Detection:
BioSuite 450,
Molecular Weight
10 5
13 m SEC
10 4
Column
BioSuite 125
BioSuite 250
BioSuite 450
Globular Protein
Mol. Wt. Range
Branched
Dextrans
Linear
PEG/PEO
5,000-100,000
10,000-500,000
20,000-7,000,000
1,000-30,000
2,000-70,000
4,000-500,000
500-15,000
1,000-35,000
2,000-250,000
BioSuite 125,
10 m SEC
10 3
7.5
10 Min.
Globular
Column
Volume
Suggested Protein
Mass Load for Maximum
Protein Mol. Wt.
Range
Suggested Volume
Load for Maximum
Multicomponent
Resolution*
Multicomponent
Resolution*
Description
Matrix
Inner
Diameter
Length
Silica
Silica
Silica
4.6 mm
4.6 mm
4.6 mm
300 mm
300 mm
35 mm
4.98 mL
4.98 mL
5,000-150,000
10,000 - 500,000
Less than 40 L
Less than 80 L
186002161
186002162
186002163
Silica
Silica
Silica
Silica
7.8 mm
7.8 mm
7.8 mm
6 mm
300 mm
300 mm
300 mm
40 mm
14.33 mL
14.33 mL
14.33 mL
5,000-150,000
10,000 - 500,000
20,000-7,000,000
186002164
186002165
186002166
186002167
Silica
Silica
Silica
Silica
Silica
Silica
Silica
Silica
7.5 mm
21.5 mm
7.5 mm
21.5 mm
7.5 mm
21.5 mm
7.5 mm
21.5 mm
300 mm
300 mm
300 mm
300 mm
300 mm
300 mm
75 mm
75 mm
13.25 mL
108.9 mL
13.25 mL
108.9 mL
13.25 mL
108.9 mL
5,000-150,000
5,000-100,000
10,000 - 500,000
10,000 - 500,000
20,000-7,000,000
20,000-7,000,000
186002168
186002169
186002170
186002171
186002172
186002173
186002174
186002175
Part No.
* Using a BSA protein standard in a 50 mM phosphate buffer containing salt (either 0.1 M NaCl or 0.1 M Na2SO4 ) eluent.
Useful protein mass loads will vary depending upon separation eluent, complexity of sample, and on the type of proteins contained in mixture.
In general, maximum component resolution is obtained by injecting the smallest possible volume of a dilute protein solution.
* Note: Operating flow rates for BioSuite Ultra-High Resolution (UHR) SEC columns (4.6 mm i.d.) are from 0.1-0.4 mL/min. Use of an HPLC system
(e.g. Waters Alliance HPLC System) capable of operating at these flows is essential for optimal UHR SEC column performance.
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233
Dimensions
Protein-Pak 60
7.8 x 300 mm
Protein-Pak 125
7.8 x 300 mm
Protein-Pak 300SW
7.5 x 300 mm
Protein-Pak 125 Sentry Guard Column
3.9 x 20 mm, 2/pkg (requires holder)
Sentry Universal Guard Column Holder
Glass Column
Protein-Pak 200SW
8 x 300 mm
Protein-Pak 300SW
8 x 300 mm
Inquire for additional offerings including prep.
Protein KW-803
25 mM Sodium Phosphate pH 6.8
0.72 mL/min
280 nm
1. Blue Dextran
2. Ferritin
3. Aldolase
4. Bovine Serum Albumin
5. Ovalbumin
6. Chymotrypsinogen
7. Cytochrome c
8. Cytidine
7
6
3
4 5
25 Min.
This gel filtration separation of protein standards demonstrates the ability to separate proteins in a wide range of
molecular weights in minutes for high sensitivity analysis or protein isolation up to the milligram scale.
Part No.
1,000-20,000
2,000-80,000
10,000-300,000
WAT085250
WAT084601
WAT080013
186000926
WAT046910
500-60,000
10,000-300,000
Column
Dimensions
Protein KW-802.5
8 x 300 mm
Protein KW-803
8 x 300 mm
Protein KW-804
8 x 300 mm
Protein-Pak 125 Sentry Guard Column
3.9 x 20 mm, 2/pkg (requires holder)
Sentry Universal Guard Column Holder
Particle Size
MW Range
Part No.
7 m
7 m
7 m
100-50,000
100-150,000
500-600,000
WAT035943
WAT035946
WAT036613
186000926
WAT046910
WAT011786
WAT011787
Pore Size
103
5x
8 x 104
4 x 105
1 x 106
7 x 106
7 x 106
5 x 103
N/A
N/A
Part No.
Ultrahydrogel 120
Ultrahydrogel 250
Ultrahydrogel 500
Ultrahydrogel 1000
Ultrahydrogel 2000
Ultrahydrogel Linear
Ultrahydrogel DP*
Ultrahydrogel Guard Column
Ultrahydrogel Guard Column DP*
234
120
250
500
1000
2000
Blend
120
N/A
N/A
Exclusion Limit
WAT011520
WAT011525
WAT011530
WAT011535
WAT011540
WAT011545
WAT011550
WAT011565
WAT011570
www.waters.com
Injection Volume:
Temperature:
Detection:
Instrument:
Analytes:
20 L
40 C
UV @ 220 nm
Alliance 2790, 996 PDA
1. RNAse
2. BSA
3. B-Lactoglobulin
4. Ovalbumin
Competitive 300
C18 column
P = 90.09
4
2
10
12
Symmetry300 C4 Columns
Symmetry300
C4 column
P = 138.56
14 Min.
Inner
Description
Diameter
Particle
Length
Size
Part No.
Symmetry300 C4
Symmetry300 C4
Symmetry300 C4
Symmetry300 C4
Symmetry300 C4
Symmetry300 C4
Symmetry300 C4
Symmetry300 C4
Symmetry300 C4
Symmetry300 C4
Symmetry300 C4
Symmetry300 C4
Symmetry300 C4
1.0 mm
2.1 mm
2.1 mm
2.1 mm
4.6 mm
4.6 mm
4.6 mm
4.6 mm
2.1 mm
3.9 mm
4.6 mm
4.6 mm
4.6 mm
150 mm
50 mm
100 mm
150 mm
50 mm
75 mm
100 mm
150 mm
150 mm
150 mm
50 mm
150 mm
250 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
5 m
5 m
186000276
186000277
186000278
186000279
186000280
186000281
186000282
186000283
186000285
186000286
186000287
186000288
186000289
Particle
Size
Protein-Pak Affinity
40 m
Epoxy-Activated Packing
Protein-Pak Affinity
Epoxy-Activated Microcolumn 40 m
(500 mg of material in a 3 cc syringe barrel)
Pore
Size
Detection:
Protein Load:
Qty.
Part No.
500
25 g
100 g
WAT030653
WAT030654
500
10/box
WAT035955
30 Min.
www.waters.com
235
Column:
Eluent: A:
Eluent B:
Flow rate:
Gradient:
Temperature:
Detection:
Column:
Eluent: A:
Eluent B:
Flow rate:
Gradient:
Temperature:
Detection:
6
8
30 Min.
LC
HC
TIC of
Reduced IgG1
antibody
7
15
Peak No.
1
2
3
4
5
6
7
8
9
10
Identity
HC Clip1 (with glycan variants)
LC (1-2 S-S Intact)
LC
LC+1 Da
HC Clip 2 (w/o glycan variants)
HC Clip 3 (w/o glycan variants)
Antibody Half mer
HC (with glycan variants)
HC Fragment (w/o glycan variants)
HC Fragment-18 Da
(w/o glycan variants)
BioSuite pPhenyl,
1000 A, 10 m RPC
2
9
7
10
15
30 Min.
Note: The BioSuite pPhenyl, 1000 RPC columns have a higher ligand
density compared to the BioSuite Phenyl, 1000 HIC columns and
are not recommended for hydrophobic-interaction separations.
236
10
12
14
16
Description
18
20
22
24
26
28
30 Min.
Length
Part No.
Polymer
Polymer
Polymer
Polymer
4.6 mm
2.0 mm
4.6 mm
21.5 mm
35 mm
150 mm
150 mm
150 mm
186002152
186002153
186002154
186002155
Polymer
Polymer
Polymer
2.0 mm
4.6 mm
21.5 mm
75 mm
75 mm
150 mm
186002156
186002157
186002158
www.waters.com
Description
BioSuite Phenyl 10 m HIC
BioSuite Phenyl 13 m HIC
Matrix
Inner
Diameter
Length
Part No.
Polymer
Polymer
7.5 mm
21.5 mm
75 mm
150 mm
186002159
186002160
Sample:
Column:
Eluent: A:
Eluent B:
Flow Rate:
Gradient:
Temperature:
Detection:
30
60 Min.
40 mg/mL *
25 mg/mL *
Lectin
Hydrophobic-Interaction Columns
Description
Dimensions
Part No.
8 x 75 mm
WAT035520
30 Min
www.waters.com
237
Sample Preparation
Samples for AAA can be divided into two groups. W here the protein in the
sample is the ultimate object of the analysis, the sample must be hydrolyzed to release the amino acids. In contrast, where the amino acids are
measured as metabolic indicators, no hydrolysis is required. However, the
amino acids may need to be separated from other sample constituents that
could bind amino acids or otherwise interfere with the analysis.
Purified proteins and peptides are hydrolyzed with acid at high temperature,
commonly 6N HCl at 100-105 C for 24 hours. Higher temperatures for
shorter times are sometimes used for specific purposes but with reduced
recovery and increased side reactions. Vapor-phase hydrolysis is useful for
sensitive analysis of small amounts of pure proteins.
T here are artifacts of hydrolysis. T he amine, asparagine, and glutamine,
are converted to the corresponding acids. Tryptophan is completely lost in
hydrolysis, and significant amounts of the sulfur-containing amino acids
are also destroyed. Some amino acids, e.g., threonine, serine, and tyrosine
may be oxidized while the bonds between hydrophobic residues are hydrolyzed slowly, leading to underestimates in both cases. Tryptophan may be
preserved with alkaline hydrolysis or with the use of methanesulfonic acid.
T he sulfur amino acids may be quantitatively oxidized with performic acid
to the very stable methionine sulfone and cysteic acid. Oxidation degrades
other amino acids, including tyrosine, serine, and threonine, so reduction
and alkylation of cysteine residues and carefully excluding oxygen from
the hydrolysis to preserve methionine is more useful. To correct for gradual
destruction or release, data from three hydrolysis times, for example, 24,
48, and 72 hours, are extrapolated to constant values.
238
Food and feed samples for assessing nutritional content are much more
complex. Hydrolysis requires large volumes of 6N HCl to ensure both
dispersal of the sample and a sufficient excess of acid. T he sulfur-containing amino acids are particularly important in these assays because they
are growth-limiting. Performic acid oxidation is preferred for accurate
determination of methionine and cysteine in these complex samples.
T he most complete profile is obtained by analyzing the sample with and
without oxidation.
In cell culture and fermentations, free amino acids are supplied in the media
as nutrients. A larger group of amino acids, including especially glutamine
and tryptophan must be included in the measurement for assessing the
health of the cell culture and supporting a feeding schedule.
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AccQTag Ultra
239
AMQ
Hydrolysates
Lle
Leu
Phe
Tyr
Met
Val
NVa
Deriv Peak
Cys
Pro
Ala
Thr
Glu
Asp
Arg
Gly
Ser
His
NH3
Lys
H
N
N
OH
O
O
O
AQC Reagent
1 Amino Acid:
t1/2<<1s
NH 2
OH
NH
2 Amino Acid:
t1/2<<1s
H
N
AMQ
Pro
Lle
Leu
Phe
Trp
Lys
N
O
O
HO
Cell Culture
Tyr
Met
Val
NVa
Deriv Peak
Cys
ABAA
Hy Lys 1
Hy Lys 2
Ala
GABA
Thr
Glu
Asp
Ser
Gln
Arg
Gly
AE
Tau
Orn
H2O
t1/2~15s
NH3
Hy Pro
His
Asn
NH
O
OH
H
N
N-Hydroxy
Succinimide
+ CO2
H
N
H
N
NH2
N
6-Aminoquinoline (AMQ)
3.00
4.50
6.00
7.50 Min.
The UPLC Amino Acid Analysis solution can be used for the analysis of protein hydrolysates or for
the larger set of amino acids found in cell culture media. The only differences in the methods are a
different dilution of the supplied eluent A concentrate and operation at a different temperature.
No method development or buffer reformulation is required.
176001279
176001235
186003836
186003837
186003838
186003839
WAT007571
WAT088122
186000384C
* This kit is intended to enable existing ACQUITY UPLC systems for AAA applications.
The Add on Kit contains the AccQTag Ultra Chemistries and Column, Documentation
and additional hardware accessories needed for AAA applications.
** The Refill Kit is intended to recharge the AccQTag Ultra chemistries that are a part of
the Application Add on Kit. This kit should not be purchased as part of an initial system.
240
6-aminoquinolyl-N-hydroxysuccinimidyl carbamate
US Patent 5,296,599 and European
Patent EP 0 533 200 B1
Reacts rapidly with both primary and secondary amines
Description
Literature
References
HPLC and UPLC Amino
Acid Analysis Brochure,
Literature Reference 720001946EN
www.waters.com
19
17
18
14
15
13
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
AMQ
Asp
Ser
Glu
Gly
His
NH3
Arg
Thr
Ala
Pro
Cys
Tyr
Val
Met
Lys
Ile
Leu
Phe
16
A. 50 pmol Standard
1
6
4 5
10
11
12
30 Min.
17
19
18
14
1
15
13
B. 1 pmol Standard
3
2
56
16
10
11
12
Application of the AccQTag Method to the analysis of hydrolysate amino acids is illustrated.
The high purity reagents provided in the AccQTag Chemistry Package enable high sensitivity
analysis by minimizing background amino acid content.
Proteins
Peptides
Standard
Injection Volume
10 to 20 L
5 to 10 L
10 to 20 L
5 to 10 L
5 L
*Based on protein average molecular weight=25,000, and peptide average molecular weight=1,000.
Dimensions/Qty.
Part No.
WAT052875
WAT088122
WAT052874
WAT052880
WAT052885
WAT052890
WAT052895
www.waters.com
241
Lys
Leu
Reagent
Ala
Pro
Gln
Val
R
N = C = S + NH2
CH
S
COOH
NH
NH
CH
Phe
Orn
Ile
Cys
Met
1 Me His
NH3
Cit
Asn
His
Glu
T hr
Ser
Arg
Tau
Trp
Tyr
Gly
A widely used technique for HPLC amino acid analysis is the Waters
PicoTag method. Based on an optimized system configuration, prepackaged reagents and extensive documentation, the PicoTag method provides
a turnkey, guaranteed approach to modern HPLC amino acid analysis.
Precolumn derivatization relies on the coupling reaction of the well known
Edman Degradation, the reaction of phenylisothiocyanate (PITC) with both
primary and secondary amino acids to form phenylthiocarbamyl (PTC) derivatives. The method is applicable to any sample including protein hydrolysates, physiologic fluids, feeds, foods, and pharmaceutical preparations.
COOH
66 Min.
The derivatization of amino acids with PITC is the first step of the wellknown Edman degradation reaction. The PTC-amino acid adducts are
stable and easily separated by reversed-phase HPLC. A single product is
formed for each amino acid. Most reaction by-products and all derivatization reagents are volatile, so they may be removed from the sample
by vacuum drying.
Phe
Cys
Ile
Leu
Dimensions/Qty.
Rgt
His
Ser
Gly
Arg
Asp
Glu
T hr
Ala
Met
Tyr
Pro
0.89
2.11
2.00
2.05
Val
Lys
A. Hydrolysate Standard
B. 16 Amino Acid Peptide, 40 pmol injected
T hr
Pro
Val
Lys
Courtesy of A.S. Feste, R.W. Drummond, and S.J. Dudrich, Nutritional Support Service,
St. Luke Episcopal Hospital, Houston, Texas.
Peptide
Composition
Asp
0.99
Glu
0.94
Gly
5.14
His
0.86
Reproducible and reliable plasma amino acid profiles are obtained in 66 minutes using Waters PicoTag Method.
In this analysis, 100 L plasma was diluted with an internal standard, deproteinized by centrifugal ultrafiltration,
and derivatized. The methionine sulfone (internal standard) peak represents 25 picomoles.
A
B
12 Min.
Dimensions/Qty.
Part No.
WAT091681
3.9 x 300 mm
WAT010950
WAT010947
4 x 1 liter
4 x 1 liter
100 mL bottle
WAT010960
WAT010965
WAT088119
This 12 minute analysis using Waters PicoTag Amino Acid Analysis Method
provides identification and accurate quantitation of the amino acid composition.
242
www.waters.com
Sample:
Column:
Temperature:
Eluent A:
Eluent B:
Flow Rate:
Gradient:
Detection:
AMQ
0.20
0.18
Phe
Ile
aIle
Leu
HCys
Nva
Met
Val
Tyr
Cys
GABA
AADA
Pro
BAIB
Hyl1
Hyl2
AABA
Ala
B-Ala
Thr
Glu
Cit
Derivatization Peak
Lys
Orn
0.10
Sar
AU
0.12
PSer
HyPro
NH3
His
Asn PEA
3MH
Tau
1MH
Ser
Gln
Carn
Arg
Ans Gly
EA
Asp
0.14
Cyst
0.16
Pro
Trp
Phe
Leu
Ile
Tyr
Met
Glu
Cit
AABA
Thr
Orn
Val
Ala
Gly
Ser
Arg
HyPro
NH3
His
Asn
3MH
Tau
0.00
Nva
Gln
0.04
0.02
Lys
Derivatization Peak
0.08
0.06
-0.02
4
10
12
14
16
18
20
22
24
26
28
30
32 min
The combination of Waters award-winning ACQUITY UPLC technology and market leading AccQTag
pre-column derivatization chemistry provides the robustness, reliability and ease of use required for
trouble-free operation in your research laboratory.
Description
Part No.
Description
Part No.
176001498
186004097
205000343
186000384C
186004095
186004099
186004090
186004092
**For use after initial installation using MassTrak AAA Solution Application Kit 176001459.
Individual components of the MassTrak AAA Solution Kit are not available separately
* For use with new installations of the MassTrak AAA Solution. Individual components of the
MassTrak AAA Solution Application Kit are not available separately
www.waters.com
243
Synthesis
60
99.5
50
40
99
.0
98
.5
98
.0
.5
30
97
20
97
.0
10
0
0
40
80
120
160
200
Length of Oligonucleotide
Isolation
A variety of methods exist for the lab-scale purification (25-500 nmole) of
oligonucleotides. T he advantages as well as disadvantages of techniques are
outlined in the table below.
Analysis
W hile polyacrylamide slab gel techniques can resolve long oligonucleotide
sequences, impurity detection of analyzed samples frequently involves
use of radio labeled nucleotides or gel staining techniques. T he advent of
capillary gel electrophoresis (CGE) with on-line detection helped overcome
some post-run detection issues. However, precise c hemical composition
determination of each separated oligonucleotide species cannot to determined using UV absorption detection. Furthermore, in some environments
the routine use of CGE for oligonucleotide analysis has been problematic due to method robustness and ruggedness concerns. HPLC, Waters
UltraPerformance LC Technology, and Mass Spectrometry have overcome
many challenges related to the separation, quantitation, and identification
of synthetic oligonucleotides. Rugged and robust separation and analysis
methods can frequently be performed in minutes rather than in hours. In
addition, quantitation with oligonucleotide composition confirmation is
possible using MS or hyphenated-MS methods.
Advantages
Disadvantages
Polyacrylamide Gel
Electrophoresis (PAGE)
Low mass-loading capacity. Gels are typically overloaded for purification and the resolution is compromised. PAGE does not separate N+x sequences. Manual band cutting. Excision is based on markers
without detailed knowledge of target oligo retention. Samples need to be extracted from the gel and
desalted, recovery of target oligonucleotides is low. Method is laborious; it is typically utilized only
when no other technique is suitable for the task.
Ion-Exchange Liquid
Chromatography (IEX-LC)
IEX-LC is efficient only for relatively short oligos (<20-25 mers); longer oligos are poorly resolved.
Sample is contaminated with high concentration of salts; further desalting is required. IEX columns
packed with non-porous sorbent offer improved resolution, but suffer with low mass load capacity.
When loading exceeds 10-20 nmoles (for 4.6 mm i.d. columns), the resolution is compromised. IEX-LC
does not separate N+x sequences.
Elegant, fast, and universal method for oligos of various length and
sequence. RP columns used with this method have sufficient mass
load capacity.
Does not adequately remove mismatch failure sequences (similarly as the target oligo they carry DMT
group). DMT group is labile, part of the product may be lost due to the spontaneous detritylation. DMT
residue and remaining acid have to removed after the detritylation.
Method requires efficient columns packed with small particle size sorbent. Oligonucleotide retention
and resolution partially depends on the sequence. Method development for different oligonucleotide
sequence and length probes is necessary.
244
www.waters.com
Exceptional Resolution of
Oligonucleotide Mixtures
ACQUIT Y UPLC OST C18, 1.7 m (designed or use with an ACQUIT Y
UPLC System) and XBridge OST C18, 2.5 m columns are well suited for
the analysis of detritylated oligonucleotides using ion-pair, reversedphase c hromatography. As indicated (see figure on right), separations
are comparable to that obtained by capillary gel electrophoresis (CGE)
in terms of component resolution yet analyses times are significantly
decreased using Waters UPLC Tec hnology. T he ability to resolve large
oligonucleotide sequences (e.g., N from N-1) is possible due to the
enhanced resolving power obtained using sub-3 m, BEH Tec hnology
particles. In addition, quantitation with molecular weight c haracterization of the separated target oligonucleotide product from failure
sequences is possible using Waters OST columns with hyphenated-Mass
Spectrometry methods and MS friendly eluents.
15
UV 260 nm
12
LC system:
Column:
Mobile Phase:
Flow Rate:
Column Temp:
Gradient:
Detection:
22 Min.
25
30
20
www.waters.com
30
35
4 Min.
245
Column:
Mobile Phase:
Flow Rate:
Column Temp.:
Gradient:
Detection:
60 C
19 to 26.5.5% B in 60 minutes
UV: 260 nm,
10 scans per second
MS: 1 scan per second,
0.1 sec interscan
9-11 %
ACN in 10 min
28-39 %
ACN in 10 min
UV 260 nm
15.5-23.5 %
MeoH in 10 min
23-37 %
ACN in 10 min
MS (TIC)
12-20 %
ACN in 10 min
45 Min.
10 Min.
Waters OST columns packed with BEH Tec hnology particles have shown
remarkable column longevity, under these demanding separation conditions, while maintaining outstanding separation performance. By comparison, significantly reduced column life results when traditional silica-based
columns are used under these same demanding separation conditions.
Separation of 5-25 mer Detritylated Oligodeoxythymidine Ladder
HPLC System:
Column:
Mobile Phase:
Column temp:
Gradient:
Flow Rate:
Detection:
Injection #4
Injection #1001
LC System:
Column:
Column Temp.:
Sample Injected:
Flow Rate:
Mobile Phase A:
Mobile Phase B:
Gradient:
Detection:
TEA
20 nt
8.73 min
TEA
TIC MS:
Capillary:
Sample Cone:
Extraction Cone:
Desolvation Temp.:
Source Temp.:
Cone Gas Flow:
Desolvation Gas Flow:
U
5800
C U
680 0
m/z
U G
U
U
10 Min.
Description
ACQUITY UPLC OST C18 *
ACQUITY UPLC OST C18 *
Custom ACQUITY UPLC OST C18 *
ACQUITY UPLC OST C18 *
ACQUITY OST C18
Method Validation Kit**
XBridge OST C18
XBridge OST C18
XBridge OST C18
Custom XBridge OST C18
XBridge OST C18
Method Validation Kit**
Particle
Size
1.7 m
1.7 m
1.7 m
1.7 m
2.5 m
2.5 m
2.5 m
2.5 m
12
15
18
21
Pore
Size
Dimension
135 2.1 x 50 mm
135 2.1 x 100 mm
24
27
30 Min.
Part No.
186003949
186003950
186003951
186005516
2.1x 100 mm
186004898
135 2.1 x 50 mm
135 4.6 x 50 mm
135 10 x 50 mm
186003952
186003953
186003954
186003955
4.6 x 50 mm
186004906
246
www.waters.com
XBridge OST C18 columns are the preferred offering for detritylated oligonucleotide purifications due to the availability of column sizes designed
to meet lab-scale isolation requirements. As indicated in the table below,
the c hoice of XBridge OST C 18 column dimension and operating flow
rate depends primarily on the scale of the synthesis reaction mixture.
For example, a 4.6 x 50 mm column containing XBridge OST C 18,
2.5 m material is an excellent selection when oligonucleotide mass
loads are less than or equal to 0.2 mol. Selection of the appropriate
column size for the amount of oligonucletide sample loaded is recommended to maximize component resolution and recovery of the target
product from non-desired failure sequences.
Column Temp:
Flow Rate:
Mobile Phase A:
Mobile Phase B:
Gradient:
Detection:
Target siRNA
duplex
85 nmol
RNA
Impurities
15 nmol
siRNA
Impurities
1.5 nmol
0.15 nmol
25 Min.
Dimensions
2.1 x 50 mm
4.6 x 50 mm
10 x 50 mm
19 x 50 mm*
30 x 50 mm*
50 x 50 mm*
*
mg***
Flow Rate
0.04 moles
0.20 moles
1.00 moles
4.00 moles
9.00 moles
25.00 moles
0.2 mg
1.0 mg
4.5 mg
16.0 mg
40.0 mg
110.0 mg
0.2 mL/min
1.0 mL/min
4.5 mL/min
16.0 mL/min
40.0 mL/min
110.0 mL/min
** Values are only approximates and vary depending on oligonucleotide length, base composition,
and heart-cutting fraction collection method used.
*** Estimated for average oligonucleotide MW and synthesis yield.
Literature
References
Purified Oligonucleotide
Purity > 95%
140 nmol
N-1
Purity 78%
*
0
84 nmol
28 nmol
1.4 nmol
0
18 Min.
www.waters.com
247
Description
ACQUITY UPLC BEH300 C18
Dimension
Particle Size
Part No.
2.1 x 50 mm
1.7 m
186003685
Waters ACQUITY
ACQUITY UPLC PST BEH300 C18 , 2.1 x 50 mm, 1.7m
50 C
0.2 mL/min
0.1M TEAA, pH 7.0
20% ACN in A
57.5 84.5 % B in 20.0 min
PDA, 260 nm
20 Min.
Gradient:
Flow Rate:
Temperature:
1. 13 bp
2. 121 bp
3. 383 bp
4. 929 bp
5. 1060 bp
6. 1857 bp
45
3
2
1
35 Min.
Absorbance 260 nm
0.040
Description
0.000
0
248
15
30 Min.
Dimensions
Part No.
4.6 x 100 mm
WAT015490
www.waters.com
15 nt
oligodeoxythymidine fragments
20 nt
25 nt
30 nt
35 nt
Qty.
Part No.
1/pk
186004135
10 Min.
Waters ACQUITY UPLC analysis of MassPREP OST Standard on an ACQUITY UPLC OST C18, 1.7 m column.
The main components are labeled. Small peaks eluting between labeled oligonucleotides are N-1, N-2, etc.
failure sequences generated during the oligonucleotide syntheses. The ACQUITY UPLC system is equipped
with 50 L standard mixer and PDA detector (260 nm).
High sensitivity
Sample concentrating
High throughput
900
800
Matrix Solution:
700
Mode:
Abundance
600
500
400
300
200
100
0
4000
4500
5000
5500
M/Z
Matrix Solution:
1600
Mode:
1400
1200
Abundance
1000
800
600
400
Description
Part No.
200
0
4000
www.waters.com
186001828BA
4500
5000
5500
M/Z
249
GLYCaN aNaLYSIS
ACQUITY UPLC BEH Glycan Column Separation of 2-AB Labeled Human IgG Glycans
UPLC Conditions
Sample:
Sample Concentration:
Sample Volume:
Injection Mode:
Column:
Eluent A:
Eluent B:
Temperature:
Detection:
Gradient:
Instrument Conf.:
Time
Flow rate
(min)
(mL/min) % A
%B
Init
0.5
25
75
46.5
0.5
40
60
48
0.25
100
0
49
0.25
100
0
50
0.5
25
75
63
0.5
25
75
Mixer (Part No. 205000403).
Needle (Part No. 205000507).
No in-line filter. No VanGuard column.
1. 2AB
Curve
6
6
6
6
6
6
6,7. 2AB
2. 2AB
8. 2AB
13. 2AB
3. 2AB
9. 2AB
4. 2AB
12. 2AB
10. 2AB
5. 2AB
11. 2AB
N-Acetylglucosamine
Fucose
Manose
Sialic Acid
Galactose
10
The N-linked glycans from pooled human IgG contain high mannose as
well as neutral, and sialylated complex structures. The chromatogram
shows 35 minutes of a one hour analysis. This challenging sample
includes high mannose, bisecting GlcNAc and sialylated species.
13
3
4
7
5
8
9
10
250
15
Glycan Analysis
20
12
11
25
30
35 min
www.waters.com
GLYCaN aNaLYSIS
High Quality Separations of Varied Glycan Structures
glycan. Waters ACQUITY UPLC BEH Glycan column was specifically
designed for use on Waters ACQUITY UPLC System to give improved
component resolution compared to traditional LC techniques for more
certain identification and reliable quantitation.
Several complementary analytical techniques are routinely used to characterize, identify, and quantitate oligosaccharides isolated from glycoproteins.
Hydrophilic-interaction liquid chromatography (HILIC) is a well-recognized
and reliable technique that can effectively separate fluorescently-labeled
6
2
10
11
12
13
14
15
16
17
18
19
20
Curve
6
6
6
6
6
6
10
15
20
25
30
35
40
21
22
45 min
The fast, high-resolution separation of a homopolymeric series of glucose oligomers illustrates the wide range of glycan sizes, extending beyond 22 glucose units that can be separated with the
ACQUITY UPLC BEH Glycan column. These labeled glycans can provide calibration reference points that are helpful in identifying more complex glycans based on relative retention times.
ACQUITY UPLC BEH Glycan Column Separation of 2AB-Labeled High Mannose Glycans from Ribonuclease b
UPLC Conditions
Sample:
Sample Concentration:
Sample Volume:
Injection Mode:
Column:
Eluent A:
Eluent B:
Temperature:
Detection:
Gradient:
1. Mannose 5
2AB
4. Mannose 8
2AB
2. Mannose 6
2AB
5. Mannose 9
2AB
Mannose 7
isomers
2AB
3a, 3b.
2*
Curve
6
6
6
6
6
4*
3*
5
10
15
20
25
30
35 min
A series of high mannose oligosaccharides was released from bovine ribonuclease b and labeled with 2-AB. The HILIC separation
performed on ACQUITY UPLC Glycan column shows both different numbers of mannose residues and isomeric structures.
Part No.
186004740
186004741
186004742
186004739
Note: ACQUITY UPLC BEH Glycan, 1.7 m columns are designed for use with the
ACQUITY UPLC system. The benefits of the small particle packing in ACQUITY UPLC
BEH Glycan, 1.7 m columns are only realized with the low system volume and low
detector dispersion of an ACQUITY UPLC system.
186004907
www.waters.com
Glycan Analysis
251
Glycan analysis
MassPREP Glycoanalysis Kit
Protein Removal
MassPREP HILIC Elution Plate
T he MassPREP Glycoanalysis kit provides simple and robust sample preparation without compromising sample recovery. As shown (right), unlabeled
or 2-AB labeled glycans can be successully analyzed by MALDI-MS or by
LC with fluorescence detection following MassPREP Glycoanalysis Kit use.
Fucose
Manose
Galactose
A)
(M+Na)
m/ z
G0
1339.47
G0F
1485.53
G1F
1647.58
G2F
1809.63
G1F
10 0
G0 F
LC-FLR
G2F
G0
0
B)
Description
Dimensions/Qty.
Part No.
2AB - G0F
10 0
2AB - G1F
186002817
2AB - G2 F
2AB - G0
96-well
1 mg vial
186002780
186001860
252
Glycan Analysis
0
1200
1400
1600
1800
2000
2200
2400
2600
2800
m /z
MALDI-TOF MS spectra of: (A) 8 pmol of the unlabeled IgG1glycans after first HILIC cleanup prior
to labeling. (B) 3 pmol of the 2-AB-labeled IgG1 glycans after second HILIC cleanup.
Each spectrum shows well purified glycans with no inferences from impurities.
www.waters.com
LC/MS
Proteomics
Protein
Characterization/QC
RapiGest SF
RapiGest SF
MassPREP Phosphopeptide
Enrichment Kit
www.waters.com
253
* BioSuite C18 3 m, Peptide Analysis - A Column (2.1 x 150 mm). Eluent A: 0.02% TFA
in water, Eluent B: 0.016% TFA in acetonitrile. Gradient: 0-50% B in 30 minutes. Flow:
0.20 mL/min. Temp: 40 C.
** Monoisotopic molecular weight
254
Enolase T37
Enolase T35
Melittin
Peptide sequence
RGDSPASSKP
DRVYIHP
RPPGFSPFR
DRVYIHPF
DRVYIHPFHL
DRVYIHPFHLLVYS
WLTGPQLADLYHSLMK
YPIVSIEDPFAEDDWEAWSHFFK
GIGAVLKVLTTGLPALISWIKRKRQQ
RASG-1
pKa
9.34
7.35
12.00
7.35
7.51
7.61
7.34
3.97
12.06
Allantoin
Molecular weight **
(g/mol)
158.0440
1000.4938
898.4661
1059.5613
1045.5345
1295.6775
1757.9253
1871.9604
2827.2806
2845.7381
AU
Component
Peak # name
1 Allantoin (V0 marker)
2 RASG-1
3 Angiotensin frag.1-7
4 Bradykinin
5 Angiotensin II.
6 Angiotensin I.
7 Renin substrate
8 Enolase T35
9 Enolase T37
10 Melittin
Renin substrate
0.5
Bradykinin
Angiotensin II
Angiotensin I
0.0
0
10
20
30 Min.
Description
MassPREP Peptide Standards
MassPREP Peptide Standards
Qty./Pack
Part No.
1 vial
5 vials
186002337
186002338
Note: The peptide mixture contains approximately 1.5 g (~1 nmole) of each peptide
www.waters.com
Description
Qty./Pack
Part No.
186002330
186002326
186002327
186002328
186002329
186002325
Qty./Pack
Part No.
1 vial
186004900
Molecular Weight, MW
Isoelectric Point, pI
13.7 k
9.6
12.4 k
10.25
66.4k
5.8
16.7 k
6.53
46.7 k
6.53
97 k
7.18
T he MassPREP phosphopeptide standard-enolase contains four phosphopeptides based on tryptic yeast enolase peptides. T he MassPREP enolase digest
with phosphopeptide mix contains equimolar mixture of enolase digest and
four phoshopeptides.
T18 1P
900
1000
1100
T18 1P
T43 2P
T19 1P
15
T19 1P
w/enolase
peptides
10
1300
1400
1500
m/z
1600
LC MS
10
1200
T18 1P
15
www.waters.com
20
Synthetic
Phosphopeptides
Sequence
Amino Acid
Residue
[M + H]+
[M + 2H]2+
Enolase T18 1P
NV PL(pY) K
126-131
813.3912
407.1995
Enolase T19 1P
HLADL (pS)K
132-138
863.4028
432.2053
Enolase T43 1P
VNQIG (pT)LSES IK
346-357
1368.6776
684.8428
Enolase T43 2P
VNQIG TL(pS)E(pS) IK
346-357
1448.6439
724.8259
T hese standards can be used on many Waters instrument systems, including the MALDI micro
MX, ACQUITY UPLC, Alliance , ZQ, Q-Tof, LCT Premier, and nanoACQUITY UPLC systems. Either
UV or MS detection is possible.
T43 1P
25
30
35
40
45 Min.
Description
T43 2P
20
25
30
35
40
45 Min.
Qty./Pack
Part No.
186003285
186003286
186003287
186003285
186002325
255
MPDS Mix 1 and MPDS Mix 2 Separation Using ACQUITY UPLC BEH
C18, 1.7 m, 75 m x 100 mm nanoACQUITY UPLC Column
MPDS Mix 2
MPDS Mix 1
MPDS Mix 2
[Molar ratio, Amount (pmol)]
ADH
1.0, 50 pmol
1.0, 50 pmol
GPB
1.0, 50 pmol
0.5, 25 pmol
ENO
1.0, 50 pmol
BSA
1.0, 50 pmol
Protein
Sample:
MPDS Mix 1: Alcohol Dehydrogenase
25 fmol/L, PhosphorylaseB 25 fmol/
L; Enolase 25 fmol/L; Bovine Serum
Albumin 25 fmol/L MPDS Mix 2: Alcohol
Dehydrogenase 25 fmol/L; Phosphorylase
B 12.5 fmol/L; Enolase 50 fmol/L;
Bovine Serum Albumin 200 fmol/L
Column:
Mobile Phase A:
Mobile Phase B:
Gradient:
Injection Volume:
Detection:
Ionization:
Mode:
Description
Qty.
Part No.
1/pk
1/pk
186002865
186002866
15 nt
20 nt
25 nt
35 nt
256
30 nt
Qty./Pack
Part No.
1 vial
186004135
10 Min.
Waters ACQUITY UPLC analysis of MassPREP OST Standard on an ACQUITY UPLC OST C181.7 m column.
The main components are labeled. Small peaks eluting between labeled oligonucleotides are N-1, N-2, etc.
failure sequences generated during the oligonucleotide syntheses. The ACQUITY UPLC system is equipped
with 50 L standard mixer and PDA detector (260 nm).
www.waters.com
Intact Myoglobin
Asp-N
Lys-C
Glu-C
10
15
20
25
30
10
15
min
20
25
30
min
RapiGest SF
Cleavage Sites
Cleavage Sites
Deconvoluted Mass
Trypsin *
Solution
C) 0.1% RapiGest SF
800
1000
1200
1400
m/z
1600
1800
2000
42000
43000
44000
mass
45000
46000
LC/MS spectra of deglycosylated ovalbumin are shown. A) Ovalbumin was solubilzed without the use of denaturant, and was not deglycosylated. B) Ovalbumin was denatured using 0.1% n-octyl-b-glycopyranoside (OG) and
deglycosylated. C) Ovalbumin was denatrued in 0.1% RapiGest SF solution and deglycosylated. The MS scans
were deconvoluted to the MW of the protein. Complete deglycosylated was observed after 2 hours deglycosylation
for the RapiGest SF solubilized ovalbumin.
Trypsin
Activity (%) **
No additive
0.1% RapiGest
0.5% RapiGest
0.1% SDS
0.5% SDS
100
100
100
24
1
(CH2)3SO3-Na+
67
29
87
97
83
CH3(CH2)10
Low pH
CH3(CH2)10
Literature
References
www.waters.com
O
O
Trypsin
Activity (%) **
Enzyme-Friendly, Mass
Spectrometry-Compatible
Surfactant for In-Solution
Enzymatic Digestion of
Proteins, Yu YQ, Gilar M, Lee
PJ, Bouvier ES, Gebler JC,
Anal Chem. 2003 Nov 1;
75(21):6023-8.
Trypsin *
Solution
O
HO
(CH2)3SO3-Na+
OH
RapiGest hydrolyzes under acidic condition (half life time = 7.6 minutes at pH 2).
Therefore, it is compatible with LC/MS and MALDI MS analysis.
Description
Rapi Gest SF 1 mg vial
Rapi Gest SF 1 mg vial (5 pack)
Rapi Gest SF 10 mg vial
Rapi Gest SF 50 mg vial
Rapi Gest SF Custom
Part No.
186001860
186001861
186002123
186002122
186002118
257
Inject #3 = 10 L mAb
(1 g/L PBS)
Inject #2 = 10 L PBS Blank,
following a 5 g mAb injection
Qty.
Part No.
1/pk
2/pk
186004032
186002785
1/pk
176001519
1/pk
WAT097958
Inject #2 = 10 L mAb
(1 g/L PBS)
Pre-run Blank
Max:2 Counts
754 Counts
Post-run Blank
5 Counts
Combined ESI-TOF mass spectra of an intact IgG1 antibody from a 4 minutes LC/MS analysis.
The results reveal no detectable carryover following a 0.5 g injection of the antibody.
MS
Light Chain
MS
Heavy Chain
TIC
(1.3 g load)
Over a series of 100 injections, satisfactory results were obtained for BSA and a mAb,
as shown for injections #97-100 on a MassPREP on-line desalting cartridge.
Reference Desalting of Proteins Using MassPREP On-line Desalting Cartridges Prior to
Mass Spectrometry. 2005 Waters Applications Note 720001077EN
258
Total ion chromatogram (TIC) from UPLC/MS analysis of light and heavy chains
from a reduced IgG1 antibody. A 10 min LC/MS run largely resolved the earlier
eluting light chain from the later eluting glycosylated heavy chains.
www.waters.com
Column:
Trapping Columns:
Mobile Phase:
Gradient Conditions:
Flow Rate:
Injection Volume:
Temperature:
MS Conditions:
Ion Source:
Source Temperature:
Desolvation
Temperature:
Instrument:
10
12.5
15
17.5
20
Description
Symmetry300 C18 Trap Column 5/pack
Atlantis dC18 Trap Column 5/pack
SCX 300 Trap Column 5/pack
Symmetry C18 Trap Column 5/pack
0.18 i.d. Custom Trap Column 5/pack
0.50 i.d. Custom Trap Column 5/pack
22.5
25
Electrospray positive
80 C
150 C
Waters CapLC with
Q-tof micro technology
27.5
30 Min.
Particle Size
Dimension
Part No.
5 m
5 m
5 m
5 m
Custom
Custom
0.18 x 23.5 mm
0.18 x 23.5 mm
0.50 x 23.5 mm
0.18 x 23.5 mm
0.18 x 23.5 mm
0.50 x 23.5 mm
186002622
186002574
186002623
186002808
186002687
186002806
NanoEase Trap Column hardware obtained from Optimize Technologies Inc., Oregon City, OR, 97045-0009, USA
www.waters.com
259
Gradient:
Flow Rate:
System:
Eluent A:
Eluent B:
5-50% B in 60 min
400 nL/min
Waters CapLC operated in
split flow mode, Q-Tof micro
100
TOF MS ES+
TIC
9.78e4
Inj #4
100
%
Inj #3
TOF MS ES+
TIC
1.48e5
Inj #2
TOF MS ES+
TIC
1.71e5
Inj #1
TOF MS ES+
TIC
1.69e5
100
%
0
100
%
0
10
Description
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
Atlantis dC18
Atlantis dC18
Atlantis dC18
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry300 C18
Atlantis dC18
Atlantis dC18
Symmetry C18
Symmetry C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
Atlantis dC18
Atlantis dC18
Atlantis dC18
Inner Diameter
Length
Particle Size
Part No.
75 m
75 m
75 m
75 m
75 m
75 m
75 m
75 m
75 m
100 m
100 m
100 m
100 m
100 m
100 m
150 m
150 m
150 m
150 m
150 m
150 m
150 m
150 m
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
150 mm
100 mm
150 mm
50 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3 m
3 m
3 m
3.5 m
3.5 m
3.5 m
3.5 m
3 m
3 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3 m
3 m
3 m
186002188
186002189
186002190
186002191
186002192
186002193
186002194
186002195
186002197
186002201
186002202
186002203
186002206
186002208
186002209
186002459
186002461
186002462
186002463
186002464
186002466
186002467
186002468
Description
Custom
Custom
Custom
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
Symmetry300 C18
Atlantis dC18
Atlantis dC18
Atlantis dC18
Atlantis dC18
Atlantis dC18
Atlantis dC18
XBridge BEH130 C18
Custom
20
30
40
50
60 Min.
Inner Diameter
Length
Particle Size
Part No.
75 m
100 m
150 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300 m
300m
Custom
Custom
Custom
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
100 mm
150 mm
50 mm
Custom
Custom
Custom
Custom
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
3.5 m
3.5 m
3.5 m
5 m
5 m
5 m
3 m
3 m
3 m
5 m
5 m
5 m
5 m
Custom
186002213
186002214
186002472
186002581
186002582
186002583
186002584
186002585
186002586
186002587
186002588
186002589
186002590
186002591
186002592
186002593
186002594
186002595
186002596
186002597
186002598
186003682
186002605
NOTE: Use of Waters 300 m Capillary Inlet Tubing (Part Number 430001471) and 300 m Capillary Outlet Tubing (Part Number
430001472) required for use with 300 m I.D. capillary columns. Waters Kit Part Number 700002757 contains both the 300 m
Capillary Inlet Tubing (Part Number 430001471) and the 300 m Capillary Outlet Tubing (Part Number 430001472).
For Peptide Separation Technology NanoEase and Capillary Columns, See Page 207
260
www.waters.com
TOF MS ES+
TIC
7.93e3
20.02 20.48
22.44
28.21
17.59
16.41
19.18
27.62
32.36
14.73 15.63
25.54
15
20
29.46 29.85
25
30
35 Min.
The ability to operate at higher pressures enables the use of longer columns
(up to and including 250 mm in length) and further leverage the separation
power of 1.7 m particles. Thus, the nanoACQUITY UPLC chemistry
offering now includes the Peptide Separation Technology nanoACQUITY
UPLC columns, which incorporate the 1.7 m BEH130 material.
3 m conventional column
Pressure 3,500 PSI
Peak Capacity 297
1.7 m BEH
Pressure 9,500
Peak Capacity 458
Column i.d.
Flow Rate
300 m
4 L/min
150 m
1 L/min
75 m
250 nL/min
30
40
50
60
70
80
90 Min.
30
40
50
60
70
80
90 Min.
* Mixture includes:
MassPREP Enolase Digestion Standard
MassPREP Phosphorylase b Digestion Standard
MassPREP Bovine Hemoglobin Digestion Standard
MassPREP ADH Digestion Standard
MassPREP BSH Digestion Standard
Part Number
186002325
186002326
186002327
186002328
186002329
Flow Rates and Column Internal Diameters for Capillary and Nanoflow. Capillary-scale separations
are done in a flow rate range of 1 l/min to 10 l/min on columns with internal diameters (i.d.)
ranging from 300 m to 100 m. Nanoflow is <1 l/min on columns with i.d. 75 m.
www.waters.com
261
Particle
Size
Inner
Diameter
Length
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3.5 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
3 m
75 m
75 m
100 m
100 m
150 m
150 m
300 m
300 m
75 m
75 m
100 m
100 m
150 m
150 m
300 m
300 m
100 mm
150 mm
100 mm
150 mm
100 mm
150 mm
100 mm
150 mm
100 mm
150 mm
100 mm
150 mm
100 mm
150 mm
100 mm
150 mm
186003491
186003492
186003493
186003494
186003495
186003496
186003497
186003498
186003499
186003500
186003501
186003502
186003503
186003504
186003505
186003506
180 m
23 mm
186003507
Part No.
Descript ion
nanoACQUITY UPLC
Trap, Symmetry C18,
(std fitting)
nanoACQUITY UPLC
Trap Symmetry C18,
(v/v fitting)
Custom
Particle
Size
Inner
Diameter
Length
Part No.
5m
180 m
20 mm
186003514
5m
180m
20mm
186004630
186003513
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
1.7 m
75 m
75 m
75 m
75 m
100 m
150 m
75 m
100 m
150 m
100 mm
150 mm
200 mm
250 mm
100 mm
100 mm
100 mm
100 mm
100 mm
186003542
186003543
186003544
186003545
186003546
186003550
186004639
186004640
186004641
For use with nanoACQUITY UPLC systems rated to 10,000 psi only.Not for use with nanoACQUITY UPLC systems rated to 5,000 psi.
Part No.
262
176001367
430001575
Part No.
430001576
430001577
430001629
186003507
186003514
186003542
186002330
715001357
Qty.
Stapels, M and Fadgen, K (2009) A Reproducible Online 2D Reversed Phase High-Low pH Method
for Qualitative and Quantitative Proteomics Current Trends in Mass Spectrometry
www.waters.com
A) Control
T18_1P
T19_1P
Enolase peptides :
4 phosphopeptide stdandard
in 1:1 molar ratio
Easy to use
T43_1P
T43_2P
B) After Enrichment
No Non-Specific
Binding
MassPREP Phosphopeptide Enrichment Kit, developed for selective enrichment of phosphopeptides from complex samples, includes a 96-well microscale, solid-phase extraction (SPE) plate packed with an affinity sorbent.
T he kit also includes a unique chemical (Enhancer) that can be added to
further improve the selectivity of phosphopeptides. In contrast to currently
popular immobilized metal affinity chromatography (IMAC) technology,
this isolation method does not require pre-loading the sorbent with metal
chelator. T his phosphopeptide enrichment protocol is simplified. T he robustness of this method is provided by the sorbent that has high native affinity
towards phosphopeptides and the Enhancer which yields superior selectivity
of the sorbent towards phosphopeptides.
20
30
Description
40
50 Min.
Units/Kit Amt/Unit
Part No.
186003864
1 plate
1 nmole
500 mg
500 mg
186003820
186003286
186003821
186003863
Microscale Kit
Nanoscale Kit
Description
Microscale Kit
Kit Contains :
nanoACQUITY UPLC 300 m x 150 mm
Atlantis dC18 3 m Column
MPDS Mix 1
MPDS Mix 2
MPDS Enolase
RapiGest SF, 1 mg
www.waters.com
Qty.
Part No.
186003903
1
1
1
1
1
186003506
186002865
186002866
186002325
186001860
Description
Qty.
Nanoscale Kit
Kit Contains :
nanoACQUITY UPLC 75 m x 150 mm
Atlantis dC18 3 m Column
nanoACQUITY UPLC 180 m x 20 mm
Symmetry C18 5 m Trap Column
MPDS Mix 1
MPDS Mix 2
MPDS Enolase
RapiGest SF, 1 mg
Part No.
186003904
Kits
186003500
1
1
1
1
1
186003514
186002865
186002866
186002325
186001860
263
Waters offers a collection of powerful tools for c haracterizing biopharmaceuticals. T hese solutions are designed specifically to provide more
accurate, reliable, and robust results, simplify your analyses and thereby
increase productivity. Our system solutions combine Waters market-leading
instrumentation, column chemistries, and software and informatics products,
that leverage the sensitivity and resolution of Ultra Performance LC
(UPLC) Tec hnology.
UPLC Technology was introduced by Waters as a novel chromatographic
technique that takes full advantage of sub-2 m column chemistries used
in a system optimized for pressures necessary to produce best-in-class
separations. By packing columns with very small particles, band-broadening
during separation is minimized. In addition, since distances are reduced in
small particles, separations can be optimized using higher linear velocities
(i.e., flow rates), while maintaining chromatographic equilibrium. T he result
of these chromatographic developments is separations with dramatically
increased resolution, sensitivity, and speed.
T he properties of these columns places a new level of demand on the
c hromatographic system, and Waters has holistically developed a family
of U PLC instruments, including t he binary-based ACQUIT Y U PLC,
nanoACQUITY UPLC, and PAT ROL UPLC systems and the quaternary-based
ACQUITY UPLC H-Class and ACQUITY UPLC H-Class Bio systems, to take
advantage of these chromatographic principles.
264
To gain the benefits of the small particles columns, the system must
generate exceptionally accurate and precise gradients at relatively high
operation pressures. T he sample manager must provide accurate, smallvolume injections with minimum cycle time. Furthermore, the detectors
must truly minimize band-broadening while providing high signal-tonoise ratios for peaks that are narrow in both time and volume.
Based on chromatographic principles, Waters synergistically developed the
columns and the instrumentation that combine to make UPLC technology a
realityand now a proven success in laboratories around the world. W hile
the first successful application of UPLC technology was in small molecule
pharmaceuticals, Waters dedication to developing impactful laboratory
innovations quickly saw the benefits of UPLC tec hnology applied to the
separation of biological and bioc hemically-significant macromolecules.
U PLC tec hnology brings a new level of separation and detection
performance to the analysis of biomolecules, whether from complex
peptide mixtures or from a potentially life saving monoclonal antibody
biotherapeutic. Today, Waters UPLC technology provides improvements
in the resolution, sensitivity, separation, and detection efficiency for
peptides,proteins, amino acids, synthetic DNA and RNA oligonucleotides,
and glycans t hat help scientists and c hromatographers ac hieve
desired objectives.
www.waters.com
Amino Acid
Analysis
Protein
Analysis
www.waters.com
Synthetic
Oligonucleotides
Analysis
Glycan
Analysis
265
267
Introduction
268
Selecting a Column
For over 40 years, Waters has been the market leader in GPC analysis,
providing the highest quality GPC products and expert applications support.
To solve unique separation problems across a broad spectrum of applications, we manufacture a wide range of GPC columns and accessories. Our
series of Ultrastyragel and Styragel columns have been the standard
in GPC columns for the separation of organic-soluble samples. To solve
your specific problems in aqueous separations, you can select from our
Ultra hydrogel columns.
Calibration Standards
Waters offers calibration standards for organic and aqueous GPC/SEC.
Standards are available as individual molecular weights, or come in kits
containing a range of MW standards.
www.waters.com
www.waters.com
269
101
102
103
104
105
106
107
108
Applications:
UHDPE
UH-Polystyrene
UH-Isoprene
UH-PMMA
Styragel HMW 6E
Styragel HMW 2
Styragel HT 6E
Styragel HT 6
Applications:
ABS
Acetyl
Acrylics
EVA
Nylon
PEEK
PET/PBT
Styragel HT 5
Styragel HT 4
Styragel HT 3
Styragel HT 2
Styragel HR 5E
Styragel HR 4E
Styragel HR 6
Applications:
Additives
Epoxy
Oligomers
Phenalics
Unsaturated Polyester
Urea/Formaldehyde
Polyethylene Glycol
Ethanolamines
Melamine Resin
Styragel HR 5
Styragel HR 4
Styragel HR 3
Styragel HR 2
Styragel HR 1
Styragel HR 0.5
270
101
102
103
104
105
106
107
108
www.waters.com
www.waters.com
Polymer
GPC Solvent
Polyisobutylene
Toluene
Polybutylene
Chlorinated Rubber
Polybutadiene
Polyisoprene
Polydimethylsiloxane
Toluene/75%
Chlorinated Polyethylene
Polyethylene Ethylacrylate
Polyethylene Vinylacetone
Polyethylene Methacrylic acid
Polyphenyleneoxide
Poly-4-methylpentene(1)
Polyethylene
TCB/135 - 160 C
TCB/135 - 160 C
Polyetheretherketone
Polyetherketone
Phenol/TCB 1:1/145 C
Polycarbonate
Methyl Chloride
Polyglycolic acid
gamma-Butyl Lactone
Acrylonitrile Methylmethacrylate
Cellulose Acetate
Cellulose Acetate Butyrate
Cellulose Acetate Proprionate
Cellulose Nitrate
Cellulose Proprionate
Cellulose Triacetate
Diallyl Phthalate
Ethyl Cellulose
Epoxy
Polyester Alkyd
Polybutene(1)
Polybutadiene Styrene
Phenol Formaldehyde
Phenol Furfural
Polymethylmethacrylate
Polypropyleneglycol
Polystyrene
Polysulfone
Polyvinylacetate
Polyvinylbutyral
Polyvinylchloride
Polyvinylchloride Acetate
Polyvinyldienechloride
Polyvinylformal
Polystyrene Acrylonitrile
Polystyrene Alphamethylstyrene
Polyester T hermoset
Phenolics
Rosin Acids
Polyglycolic Acid
THF/40 C
Melamine Formaldehyde
Nylon (All types)
Polybutylene Terphthalate
Polyethylene Terphthalate
Hexafluoroisopropanol + 0.075 M
Sodium Trifluoroacetate/55 C
or
m-Cresol + 0.05 m LiBr/100 C
Poly Acrylonitrile
ABS (Acrylonitrile Butadiene Styrene)
ASA (Acrylic Styrene Acrylonitrile
ABA (Acrylonitrile Butdiene Acrylate)
Carboxymethyl Cellulose
ABS/Polycarbonate
Polybutadiene Acrylonitrile
Polyurethane
Polyacetal
Polyoxymethylene
Polyimide
Polyamide imide
Polyetherimide
Polyethersulfone
Polyvinyldienefluoride
N-Methyl Pyrrolidone
+ 0.05 m LiBr/100 C
Polyfuran Formaldehyde
Dimethylacetamide/60 C
Shipping Solvent
271
Conditions:
Sample:
Mobile Phase:
Flow Rate:
Column Bank:
Mobile Phase:
Temperature:
Flow Rate:
Sample:
Polystyrene
THF
1 mL/min
Styragel HR 0.5
Styragel HR 1
Styragel HR 2
Styragel HR 3
Styragel HR 4
Styragel HR 5
Styragel HR 6
Styragel HR 4E
Styragel HR 5E
20
25
30
35
40
45 Min
20
25
30
35
40
45 Min
Part No.
Weight Range
Part No.
THF
Part No.
DMF
Toluene
0-1,000
100-5,000
500-20,000
500-30,000
5,000-600,000
50-100,000
50,000-4 x 106
2,000-4 x 106
200,000-1 x 107
WAT044231
WAT044234
WAT044237
WAT044222
WAT044225
WAT044240
WAT054460
WAT044228
WAT054468
WAT044232
WAT044235
WAT044238
WAT044223
WAT044226
WAT044241
WAT054466
WAT044229
WAT054474
WAT044230
WAT044233
WAT044236
WAT044221
WAT044224
WAT044239
WAT054464
WAT044227
WAT054470
WAT054405
WAT054415
WAT054410
272
Part No.
Weight Range
Part No.
THF
Part No.
DMF
Toluene
0-1,000
100-5,000
500-20,000
500-30,000
5,000-600,000
50-100,000
2,000-4 x 106
WAT045835
WAT045850
WAT045865
WAT045880
WAT045895
WAT045805
WAT045820
WAT045840
WAT045855
WAT045870
WAT045885
WAT045900
WAT045810
WAT045825
WAT045830
WAT045845
WAT045860
WAT045875
WAT045890
WAT045800
WAT045815
www.waters.com
Conditions:
Sample:
Mobile Phase:
Flow Rate:
Polystyrene
THF
1 mL/min
Molecular Weight
Styragel HT 2
Styragel HT 5
Styragel HT 3
Styragel HT 6
Styragel HT 4
Styragel HT 6E
Styragel HT 6E
TCB
140 C
LDPE
Part No.
Weight Range
Part No.
THF
Part No.
DMF
Toluene
100-10,000
500-30,000
5,000-600,000
50,000-4 x 106
200,000-1 x 107
5,000 - 1 x 107
WAT054475
WAT044207
WAT044210
WAT044213
WAT044216
WAT044219
WAT054480
WAT044208
WAT044211
WAT044214
WAT044217
WAT044220
WAT054476
WAT044206
WAT044209
WAT044212
WAT044215
WAT044218
WAT054405
WAT054415
WAT054410
Effective Molecular
Column
Styragel HT 3
Styragel HT 4
Styragel HT 5
Styragel HT 6
Styragel HT 6E
www.waters.com
Part No.
Weight Range
Part No.
THF
Part No.
DMF
Toluene
500-30,000
5,000-600,000
50,000-4 x 106
200,000-1 x 107
5,000-1 x 107
WAT045920
WAT045935
WAT045950
WAT045965
WAT045980
WAT045925
WAT045940
WAT045955
WAT045970
WAT045985
WAT045915
WAT045930
WAT045945
WAT045960
WAT045975
273
Conditions:
Sample:
Mobile Phase:
Flow Rate:
Column Bank:
Column Dimensions:
Polystyrene Stds:
Flow Rate:
Temperature:
Solvent:
Polystyrene
THF
1 mL/min
Column Bank:
Molecular Weight
Column Dimensions:
Polystyrene Stds:
Styragel HMW 2
Styragel HMW 6E
Flow Rate:
Temperature:
Solvent:
Styragel HMW 7
Part No.
Weight Range
Part No.
THF
Part No.
DMF
Toluene
100-10,000
500,000-1 x 108
5,000-1 x 107
WAT054488
WAT044201
WAT044204
WAT054494
WAT044202
WAT044205
WAT054490
WAT044200
WAT044203
WAT054405
WAT054415
WAT054410
Part No.
THF
Part No.
DMF
Toluene
WAT046805
WAT046820
WAT046810
WAT046825
WAT046800
WAT046815
Part No.
Weight Range
108
500,000-1 x
5,000-1 x 107
274
www.waters.com
Name
Solvent
Particle Size
MW Range
Part No.
THF
THF
THF
THF
THF
THF
THF
3 m
3 m
3 m
3 m
3 m
3 m
3,5 m
100-1,000
500-10,000
1,000-20,000
2,000-60,000
10,000-400,000
500-700,000
1,000-4,000,000
186001741
186001742
186001743
186001744
186001745
186001746
186001747
THF
THF
THF
THF
THF
THF
THF
THF
THF
THF
THF
3 m
3 m
3 m
3 m
3 m
3 m
5 m
3 m
3 m
3 m
3,5 m
100-1,000
500-10,000
1,000-20,000
2,000-60,000
10,000-400,000
25,000-4,000,000
50,000-10,000,000
100-10,000
500-400,000
1,000-4,000,000
5,000->10,000,000
186001749
186001750
186001751
186001752
186001753
186001754
186001755
186001757
186001758
186001759
186001760
Water
Water
Water
Water
Water
Water
4 m
4 m
6 m
7 m
9 m
9 m
500-2,000
1,000-60,000
10,000-400,000
50,000-4,000,000
100,000->10,000,000
500-10,000,000
186001785
186001786
186001787
186001788
186001789
186001790
Aqueous GPC
HSPgel AQ 2.5
HSPgel AQ 3.0
HSPgel AQ 4.0
HSPgel AQ 5.0
HSPgel AQ 6.0
HSPgel AQ MB-H
Solvent
Particle Size
MW Range
Part No.
THF
THF
THF
THF
THF
THF
THF
THF
THF
THF
THF
THF
ODCB
ODCB
ODCB
ODCB
ODCB
ODCB
ODCB
ODCB
ODCB
ODCB
ODCB
ODCB
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
5 m
100-1,000
500-10,000
1,000-20,000
2,000-60,000
10,000-400,000
25,000-4,000,000
50,000-10,000,000
100,000->15,000,000
100-1,000
500-400,000
1,000-4,000,000
5,000->10,000,000
100-1,000
500-10,000
1,000-20,000
2,000-60,000
10,000-400,000
25,000-4,000,000
50,000-10,000,000
100,000->15,000,000
100-1,000
500-400,000
1,000-4,000,000
5,000->10,000,000
186001761
186001762
186001763
186001764
186001765
186001766
186001767
186001768
186001769
186001770
186001771
186001772
186001773
186001774
186001775
186001776
186001777
186001778
186001779
186001780
186001781
186001782
186001783
186001784
High-Temperature GPC
Room-Temperature GPC
HSPgel RT 1.0
HSPgel RT 2.0
HSPgel RT 2.5
HSPgel RT 3.0
HSPgel RT 4.0
HSPgel RT 5.0
HSPgel RT 6.0
HSPgel RT MB-L
HSPgel RT MB-L/M
HSPgel RT MB-M
HSPgel RT MB-H
Name
HSPgel HT 1.0
HSPgel HT 2.0
HSPgel HT 2.5
HSPgel HT 3.0
HSPgel HT 4.0
HSPgel HT 5.0
HSPgel HT 6.0
HSPgel HT 7.0
HSPgel HT MB-L
HSPgel HT MB-L/M
HSPgel HT MB-M
HSPgel HT MB-H
HSPgel HT 1.0
HSPgel HT 2.0
HSPgel HT 2.5
HSPgel HT 3.0
HSPgel HT 4.0
HSPgel HT 5.0
HSPgel HT 6.0
HSPgel HT 7.0
HSPgel HT MB-L
HSPgel HT MB-L/M
HSPgel HT MB-M
HSPgel HT MB-H
All columns are 6.0 x 150 mm
www.waters.com
HR
High Resolution
MB
Mixed Bed
L/M
Low/Medium MW Range
RT
Room Temperature
Low MW Range
High MW Range
AQ
Aqueous
Medium MW Range
275
KF-800 (THF)
KF-801
KF-802
KF-802.5
KF-803
KF-804
KF-805
KF-807
KF-806M (linear)
KF-G pre-column (4.6 x 10 mm)
Part No.
1.5 x 103
5 x 103
2 x 104
7 x 104
4 x 105
4 x 106
(2 x 108)
(4 x 107)
WAT030697
WAT030698
WAT030699
WAT034100
WAT034101
WAT034102
WAT034104
WAT034105
WAT034106
2 x 104
7 x 104
4 x 105
4 x 106
WAT034109
WAT034110
WAT034111
WAT034112
WAT035524
2.5 x 103
5 x 103
2 x 104
7 x 104
4 x 105
(4 x 107)
(2 x 108)
(4 x 107)
WAT034116
WAT034117
WAT034118
WAT034119
WAT034120
WAT034122
WAT034123
WAT034124
WAT034125
K-800 (Chloroform)
HFIP-803
HFIP-806M (linear)
HFP-LG precolumn (8 x 50 mm)
104
7x
(4 x 107)
Part No.
WAT035605
WAT035611
WAT035612
K-802.5
K-803
K-804
K-805
K-G precolumn (4.6 x 10 mm)
KD-800 (DMF)
KD-801
KD-802
KD-802.5
KD-803
KD-804
KD-806
KD-807
KD-806M (linear)
KD-G prec-olumn (4.6 x 10 mm)
276
www.waters.com
Column Optimization
Column:
Sample:
Solvent:
Flow Rate:
Detection:
Dimensions
Part No.
19 x 150 mm
19 x 300 mm
4.6 x 30 mm
WAT036555
WAT036554
186001913
Dimensions
Part No.
19 x 150 mm
19 x 300 mm
4.6 x 30 mm
186001915
186001916
186001914
Effective Molecular
Weight Range
Flow Rate
mL/Min
Part No.
Toluene
Part No.
THF
50-1,500
100-10,000
200-30,000
5,000-600,000
50,000-4M
200,000-10M
2,000-4M
4-10
4-10
4-10
4-10
4-10
4-10
4-10
WAT025866
WAT025867
WAT025868
WAT025869
WAT025870
WAT025871
WAT025872
WAT025859
WAT025860
WAT025861
WAT025862
WAT025863
WAT025864
WAT025865
100
500
103
104
105
106
Linear
[Effective linear functional range: 2K-4M]
www.waters.com
277
Polymer
Eluent
Nonionic
Polyethylene Oxide
Polyethylene Glycol
Polysaccharides
Pullulans
Dextrans
Cellulosics (H2O Soluble)
Polyvinyl Alcohol
Polyacrylamide
0.10M NaNO3
Nonionic Hydrophobic
Polyvinyl Pyrrolidone
Anionic
0.10M NaNO3
Anionic Hydrophobic
Cationic
DEAE Dextran
Polyvinylamine
Polyepiamine
N-Acetylglucosamine
0.80M NaNO3
Cationic Hydrophobic
Amphoteric
278
0.10M TEA
0.10M TEA/1% HOAc
Polyethyleneimine
Poly (n-Methyl-2-Vinyl Pyridinium)
Lysozyme
Chitosan
Polylysine
Peptides
Collagen/Gelatin
www.waters.com
Ultrahydrogel Columns
Packed with hydroxylated polymethacrylate-based gel, Waters Ultrahydrogel
SEC columns are ideal for the analysis of aqueous-soluble samples, such
as oligomers; oligosaccharides; polysaccharides; and cationic, anionic, and
amphoteric polymers. Measuring 7.8 x 300 mm, these high-resolution
columns offer many advantages over conventional aqueous SEC columns,
suc h as:
Gelatin Sample
Column:
Temperature:
Eluent:
Flow rate:
Detection:
Pore Size
Exclusion Limit
Part No.
Ultrahydrogel 120
Ultrahydrogel 250
Ultrahydrogel 500
Ultrahydrogel 1000
Ultrahydrogel 2000
Ultrahydrogel Linear
Ultrahydrogel DP*
Ultrahydrogel Guard Column
Ultrahydrogel Guard Column DP*
120
250
500
1000
2000
Blend
120
N/A
N/A
5 x 103
8 x 104
4 x 105
1 x 106
7 x 106
7 x 106
5 x 103
N/A
N/A
WAT011520
WAT011525
WAT011530
WAT011535
WAT011540
WAT011545
WAT011550
WAT011565
WAT011570
www.waters.com
279
Calibration Standards
Organic Calibration Standards
Description
Polystyrene Kit
Low-Mid MW*
102/10
102/10
Part No.
102/10
4x
g, 5.3 x
g, 9.5 x
g,
2.8 x 103/5 g, 6.4 x 103/5 g, 1 x 104/5 g,
1.7 x 104/5 g, 4.3 x 104/5 g,
1.1 x 105/5 g, 1.8 x 105/5 g
Polystyrene Kit
4.3 x 105/5 g, 7.8 x 105/5 g, 1.3 x 106/1 g,
Mid-High MW
2.8 x 106/1 g, 3.6 x 106/1 g, 4.3 x 106/1 g,
5.2 x 106/1 g, 6.2 x 106/1 g,
8.4 x 106/1 g, 2 x 107/1 g
Polystyrene Kit
5.8 x 102, 9.5 x 102, 1.2 x 103, 1.8 x 103,
SL-105
2.47 x 103, 3.77 x 103, 5.1 x 103,
7.6 x 103, 1.25 x 104, 1.7 x 104
Polystyrene Kit
1.2 x 103, 3.25 x 103, 1.02 x 104, 2.8 x 104,
SM-105
6.8 x 104, 1.95 x 105, 4.9 x 105, 1.08 x 106,
1.75 x 106, 2.75 x 106
Polystyrene Kit
4.5 x 105, 1.27 x 106, 2.3 x 106, 3.26 x 106,
SH-75
4.34 x 106, 8 x 106, 1.5 x 107
Polymethylmethacrylate
2.4 x 103, 9.5 x 103, 3.1 x 104,
Mid MW Kit
5.2 x 104, 1 x 105, 1.7 x 105, 2.7 x 105,
4.91 x 105, 7.3 x 105, 1 x 106
Polymethylmethacrylate
1 x 103, 1.7 x 103, 2.5 x 103,
Low MW Kit
3.5 x 103, 5 x 103, 7 x 103, 1 x 104,
1.3 x 104, 2 x 104, 3 x 104
Polybutadiene Kit
1 x 103, 3 x 103, 7 x 103, 1 x 104, 3 x 104,
7 x 104, 1 x 105, 3 x 105, 7 x 105, 1.1 x 106
Polyisoprene Kit
1 x 103, 3 x 103, 1 x 104, 3 x 104, 7 x 104,
1 x 105, 3 x 105, 5 x 105, 1 x 106, 3 x 106
WAT011588
WAT011610
500 mg/each
WAT034208
500 mg/each
WAT034209
500 mg/each
WAT034210
500 mg/each
WAT035706
500 mg/each
WAT035707
500 mg/each
WAT035709
500 mg/each
WAT035708
2.8 x 103
6.2 x 103
1.03 x 104
1.67 x 104
4.39 x 104
1.07 x 105
1.86 x 105
4.22 x 105
7.75 x 105
1.26 x 106
2.86 x 106
3.84 x 106
4.48 x 106
5.48 x 106
6.77 x 106
8.42 x 106
2.0 x 107
4 x 102
5.3 x 102
9.5 x 102
2.8 x 103
6.4 x 103
1.01 x 104
1.73 x 104
4.30 x 104
1.06 x 105
1.84 x 105
4.27 x 105
7.91 x 105
1.30 x 106
2.80 x 106
3.61 x 106
4.27 x 106
5.20 x 106
6.20 x 106
Type
ReadyCal, 4 mL
Autosampler Vial
ReadyCal, 2 mL
Autosampler Vial
Approximate
Standards
Molecular Weight
Part No.
12
4 x 102 x 2 x 106
WAT058930
12
4 x 102 x 2 x 106
WAT058931
Description
Part No.
200 mg /each
WAT034207
500 mg/each
WAT054392
500 mg/each
WAT011572
1 gram/each
WAT035711
250 mg/each
WAT035714
Part No.
10 g
10 g
10 g
5g
5g
5g
5g
5g
5g
5g
5g
5g
1g
1g
1g
1g
1g
1g
1g
1g
WAT011590
WAT011592
WAT011594
WAT011596
WAT011598
WAT011600
WAT011602
WAT011604
WAT011606
WAT011608
WAT011612
WAT011614
WAT011616
WAT011618
WAT011620
WAT011622
WAT011624
WAT011626
WAT011628
WAT011630
Qty.
Part No.
2.5 x 104
4.0 x 104
7.3 x 104
1.5 x 105
2.8 x 105
6.6 x 105
8.5 x 105
0.5 g
0.5 g
0.5 g
0.5 g
0.5 g
0.5 g
0.5 g
WAT011574
WAT011576
WAT011578
WAT011580
WAT011582
WAT011584
WAT011586
2.4 x 104
4.0 x 104
7.9 x 104
1.6 x 105
3.4 x 105
5.7 x 105
8.5 x 105
* Light scattering
* Light scattering
280
www.waters.com
HPLC Pumps
2707 Autosampler................................................302
Manual Injectors.....................................................305
282
2747 Autosampler................................................303
Detectors
2487/2488 Dual-Wavelength
Absorbance Detector .............................................306
2489 UV/Visible Dual-Wavelength
Absorbance Detector .............................................306
2996 Photodiode Array Detector .........................307
2998 Photodiode Array Detector .........................307
2414 Refractive Index Detector ...........................309
2424/2420 Evaporative Light
Scattering (ELS) Detector.......................................309
2465 Electrochemical Detector ............................309
2475 Multi-Wavelength
Fluorescence Detector ............................................310
432 Conductivity Detector ...................................310
Other Systems
Alliance GPC/V 2000 System ..............................310
Mass Spectrometers
Quadrupole Time of Flight (Q-Tof) Mass Spectrometers
Q-Tof II ....................................................................311
Q-Tof Ultima API ....................................................311
Q-Tof micro.............................................................311
Q-Tof Premier .........................................................311
Q-Tof Premier MALDI .............................................311
SYNAPT MS/HDMS .................................................311
SYNAPT G2.............................................................311
Xevo QTof ...............................................................312
Xevo G2 QTof .........................................................312
Tandem Quadrupole Mass Spectrometers
Quattro Ultima .......................................................313
Quattro Ultima Pt ..................................................313
Quattro micro .........................................................313
Quattro micro GC ...................................................313
Quattro Premier/Premier XE ..................................314
TQ Detector.............................................................314
Xevo TQ ..................................................................315
Xevo TQ-S ...............................................................315
Single Quadrupole Mass Spectrometers
EMD 1000/ZQ........................................................316
3100 Mass Detector..............................................316
SQ Detector ............................................................316
Time of Flight (TOF) Mass Spectrometers
MALDI micro MX.....................................................317
LCT...........................................................................317
LCT Premier/Premier XE .........................................317
GCT ..........................................................................318
GCT Premier .........................................................318
Magnetic Sector Mass Spectrometer
AutoSpec/AutoSpec Premier .................................318
Waters Global Services ............................................................319
General HPLC Parts
Column and Cartridge Fittings
and Accessories ...............................................................320
SLIPFREE Connectors........................................................321
PEEK Tubing and Fittings ................................................322
PTFE/ETFE Tubing .............................................................323
Stainless Steel Tubing ....................................................323
Compression Screws and Ferrules ..................................323
Filters ...............................................................................323
283
To Find Parts:
Use Our Waters Quality Parts Locator
Visit Waters online at www.waters.com/parts to use the Waters Quality Parts
Locator, which allows you to locate, view, and order instrument components.
T he Waters Quality Parts Locator offers detailed information and schematics
for parts listed in this catalog. Simply select the appropriate instrument and
click on its image to drill down to identify the part youre looking for. Youll
also find troubleshooting information from our technical experts, to help
you determine if a part should be repaired or replaced.
Literature
References
Waters Global Services: W hy Choose Waters Services Brochure
Literature Reference 720001153EN
284
Go to www.waters.com/order
Telephone:
www.waters.com
Part No.
201000174
Part No.
WAT052586
201000186
WAS081142
201000159
201000193
Part No.
i2Valve
201000197
Description
ACQUITY UPLC Binary Solvent Manager Performance Maintenance Kit
PM Kit includes: Plungers, Check valves, Seals and Filters.
Qualification/Calibration Kits
Designed to be used with Waters
Qualification Workbooks or Calibration
procedures. T hese specially-prepared
solutions come with a Material Safety
Data Sheet and a Certificate of Analysis
to assist you in successfully completing
your own qualification tasks.
For a complete product listing and pricing,
please visit Waters NEW Order Center
www.waters.com/order
www.waters.com
205000291
205000365
205000478
410001905
700001012
700003114
700003139
700003147
700003151
700003156
700003164
700003167
700003168
700003169
700003170
Part No.
700003557
700002912
700002964
700002965
700002968
430001773
430001443
WAT042091
Part No.
201000173
Part No.
201000173
205000404
205000403
430001020
430001113
430001121
430001207
430001486
700002594
700002595
700002596
700002597
700002598
700002599
700002600
700002601
700002603
700002604
700002634
700002635
700002636
700002645
700002660
700003616
700003615
700002911
700002912
700002968
WAT025531
WAT037372
WAT037373
WAT076152
285
201000174
205000343
205000362
205000363
205000369
205000370
405003740
405003741
405003742
405003743
410001347
410001659
430001084
430001086
430001221
430001264
430001311
430001320
430001325
430001326
700002569
700002570
700002572
700002644
700002765
700002775
Part No.
Flow Cell, Std., ACQUITY 2996 (For 2996 S/N K05UPD699 or lower)
Flow Cell, Low Flow., ACQUITY 2996 (For 2996 S/N K05UPD699N or lower)
Connector Plug, 10-position
Tube Assembly, Low Flow, 2996 Inlet
Tube Assembly, Std. Flow, 2996 Inlet
Capillary Tubing Cutter
PEEK Tubing Cutter
Backpressure Regulator
ACQUITY 2996 PM Kit ( For 2996 w/ S/N K05UPD699N or lower)
FUSE, 3.15A ( 250V, 5 x 20 mm), 2/pk
I/O Terminal Plug, 10-pin
Power Cord, 110V
Fuse 2.0A, 250V, Fast-Blo, 5 x 20 mm
Lamp Mounting Screw, M4 x 10
205015002
205015003
323000247
430001783
430001784
605000101
700001012
700002676
WAT052586
700001800
WAT057112
WAT097194
WAT163-14
WAT411410
Part No.
201000186
205015017
205015019
323000247
430001784
441000456
605000101
700001012
700002676
WAT097194
Part No.
205000263
205000291
205000365
425000536
700002587
700003114
700003115
700003116
Part No.
286
Description
201000186
205015000
205015001
205015016
205015009
323000247
430001749
430001748
441000456
605000101
700001012
700001800
700002676
WAS081142
700001800
WAT097194
Part No.
201000159
205000342
323000247
425000326
430001562
430001565
440000145
441000456
700002682
700002801
WAT020321
WAT057235
WAT163-18
Literature
References
Waters Educational Services Course Catalog,
Literature Reference 720000508EN
www.waters.com/courses
www.waters.com
Part No.
700003711
WAT055426
700001800
323000247
323000446
410002096
700002676
700003694
WAT057235
WAT097194
441000372
Part No.
205015017
205015019
323000247
441000456
441000373
700002676
700001800
WAT097194
WAT057235
430001784
205000505
430001783
Part No.
201000208
700003794
405003743
405003740
405003741
405003742
www.waters.com
Description
Part No.
700005269
Part No.
715002099
715002098
715002531
287
201000233
289002414
405003015
405007627
405010508
410001296
430002208
430002316
430002317
430002319
430002357
430002387
430002388
430002389
430002390
430002400
700001530
700002594
700002595
700002597
700002598
700002599
700002600
700002601
700002635
700002660
700004023
700005162
700005164
700005165
700005167
700005173
700005119
WAT076152
WAT270926
Description
ACQUITY H-Class Sample Manager- Flow Through Needle
Performance Maintenance Kit
Sample Needle Kit, 15 L
Inline Waste Valve, 1/4-28 thread
Seal Extension Tube
Injector Valve Cartridge, SM-FTN
Sample Needle Guide
Needle Seat
Syringe 100 L, HP
Part No.
201000234
700005215
410003180
700005234
700005236
405008854
405011492
700002570
Part No.
700005248
415001544
205000730
205000478
410003295
205000726
415001608
700001796
441001040
700005251
Description
ACQUITY H-Class Local Console Controller*
ACQUITY H-Class LCC Bracket*
Part No.
725000464
205000650
*Note: Both part numbers are required to properly install device to ACQUITY H-Class.
288
www.waters.com
Part No.
201000198
430001326
700002705
430001311
430002015
205000585
700004145
700004144
430001229
700004147
205000275
430001017
700002455
700004057
700002635
700002645
700002634
201000198
Literature
References
Connections INSIGHT Case Study: Reducing Instrument DownTime in an Analytical Laboratory
Literature Reference: 720001602EN
www.waters.com/insight
Description
Part No.
725000464
205000590
www.waters.com
289
Part No.
WAS081142
201000182
201000186
201000181
205000589
Part No.
201000181
289002414
289002111
289002172
430001113
430001114
430001115
430001116
430001120
430001121
430001209
430001210
430001346
430001389
430001436
430001511
430001512
430001534
430001535
430001568
430001569
430001572
700001502
700002594
700002595
700002596
700002598
700002599
WAT270868
700002600
700002601
700002603
700002604
700002635
700002645
700002660
700002696
700002702
700002712
700002713
700002968
WAT076152
WAT270868
To ensure the maximum performance and longevity of your ACQUITY UPLC System, it is critical
that you use Waters Quality Parts for maintenance. For more information on parts and accessories
for your ACQUITY & nanoACQUITY UPLC Systems, please visit www.waters.com/parts
290
www.waters.com
Part No.
201000182
205000542
205000398
289000439
289002576
289002802
405003740
405003741
405003742
405003743
405005067
405005068
410001348
430001264
430001311
430001320
430001325
430001326
430002242
430001570
430001571
430001575
430001576
430001577
430001629
700000610
700002569
700002570
700002572
700002576
700002577
700002634
700002708
700002754
700002755
700002757
700002907
700002908
Part No.
430001456
289004407
289004442
430002140
430001629
700004599
430002183
430002153
430002155
430001575
700004601
430002242
186004630
289002576
700002747
430001577
430001571
430001470
289003345
430001848
430001837
430001835
410001697
725000455
205000540
Part No.
201000186
205000159
205015013
323000247
700001800
700002676
WAS081142
WAT055426
700001800
Description
Part No.
www.waters.com
205015015
205016040
291
Description
Part No.
WAT270944
WAT045427
WAT062341
WAT062479
WAT271066
WAT271067
WAT077342
WAT077343
WAT045424
Description
Part No.
700000201
PM Kit includes: Filters and Syringe (2.5 mL), and Tubing Kit.
Parts & Accessories
HPMV tool kit
Solvent bottle caps, 4 L (4/pk)
Bottle caps, 1 L (4/pk)
Plunger seals replacement kit, black (2/pk)
Replacement plunger
Syringe, 2500 L
Syringe, 25 L
High-pressure motor valve rebuild kit
WAT045427
WAT062341
WAT062479
WAT271066
WAT271067
WAT077342
WAT077343
WAT045424
* Note: For proper maintenance of the 2690D/2695D, please make sure to order PM Kit
700000201 and PM Kit WAT270944.
292
www.waters.com
Description
Part No.
201000169
430000781
430000762
430000782
430000783
430000784
430000785
Description
Part No.
201000107
WAT077343
WAT073109
WAT077342
Syringe
2790/2795 Performance Maintenance Kit
Literature
References
Waters Educational
Services Course Catalog,
Literature Reference 720000508EN
www.waters.com/courses
www.waters.com
293
Sapphire plungers
One of the most important components is the sapphire plunger. It must be
clean and smooth if any plunger seal is expected to work for any length
of time. Waters recommends in our Performance Maintenance strategy that
you change them once each year.
Waters offers two plungers. T he standard plunger is manufactured using
the traditional processes in place by Waters and other instrument vendors.
Waters also offers the PerformancePLUS Sapphire Plunger, whic h has
the crystalline structure of the plunger oriented lengthwise, rather than
randomly. T his results in a harder surface around the circumference
of the plunger, extending its expected lifetime. T he condition of the
sapphire plungers is critical. PerformancePLUS plungers with the oriented
crystal structure will provide the best sealing surface and durability.
Plunger seals
Plunger seal material is another critical variable. Plunger seals are available in several materials that can be used for improved performance in
specific applications. Plunger seal life should typically be six months to
one year; you may try the alternative seals if the lifetime is lower.
294
Check Valves
Check valve failures can be a common cause of reproducibility problems.
T here are a number of check valve failure modes, including sticking (failure
to open or close) and intermittent leaks, which results in variable retention
times and pressure fluctuations. Intermittent leaks are often caused by
particulate matter that sticks to the c heck valve ball. T his material can
come from the mobile phase, shedded seal material, dirty glassware, or
poor laboratory cleanliness.
Waters offers optional configurations of check valve cartridges. The standard
check valve cartridge contains a synthetic ruby ball and sapphire seat.
A second option for separations modules is the PerformancePLUS Check
Valve Cartridge*. (Standard for 500 & 600 Series pumps.) T his cartridge
has a larger ball and seat (3.175 mm) and internal volume. T he ball and
seat are manufactured from ruby and sapphire. T his cartridge has been
effective in solving sticking problems and provides excellent sealing
c haracteristics. T he larger orifices in this cartridge can provide an
advantage at higher flow rates.
Another optional check valve cartridge is available with a ceramic ball and
seat which can be helpful in applications where the ruby/sapphire ball and
seat-based check valves still exhibit problems with sticking.
Check valves should perform within specification for a year or more for
most customers. T his can be greatly affected by laboratory practices,
solvent preparation, c hoice of plunger seal material, and the mobile
phase required for the application. Check valve sticking can be reduced or
eliminated by experimenting with different ball and seat sizes and materials to see which provides optimal performance for the particular application
and operating environment.
www.waters.com
HPLC Pumps
Common Parts for HPLC Pumps
2690/D
2695/D
2790
2795
2796
WAT271067
WAT270959
700001326
WAT270938
WAT271066
WAT270939
WAT270940
www.waters.com
WAT271017
WAT271018
WAT270941
700002399
700002761
700002333
700002332
WAT035190
WAT088084
WAT270927
WAT270964
700001218
WAT077343
WAT073109
700000565
700000611
WAT077342
700001247
WAT045424
WAT045427
700002791
WAT271019
WAT270328
700000383
700000384
700000385
700000389
700000564
515
205000202
205000251
205000252
205000250
WAT025296
WAT025297
WAT031790
WAT026644
700002282
WAT025384
WAT069581
WAT022934
WAT026644
WAT022946
WAT026613
WAT025296
WAT025297
700002275
WAT088223
WAT024120
WAT094003
WAT025212
WAT060307
WAT033679
WAT030541
WAT060308
WAT025203
WAT060495
WAT025028
WAT025216
WAT025207
WAT030543
WAT026014
600
1515
1525
1525
micro
Ext
Flow
700000253
700002399
700000254
WAT030852
WAS207069
WAT025656
WAT069511
WAT060304
WAT031788
WAT025360
WAT025531
WAT007272
WAT026592
WAT025746
WAT027629
WAT060305
WAT069583
626
295
Description
Part No.
WAT052587
700000253
WAT022946
WAT011042
WAT025263
WAT025297
WAT025559
WAT027629
700000254
Description
Part No.
201000113
700000254
WAT022946
WAT025531
Solvent Filter
WAT025531
296
www.waters.com
Description
Part No.
201000114
700000254
WAT022946
WAT025531
205000324
Solvent Filter
WAT025531
Description
Part No.
201000161
www.waters.com
297
Description
Part No.
201000130
700001493
700001494
Description
Part No.
201000209
205000505
410000570
430002032
430002121
700001529
700001530
700004425
700004430
700004431
700004439
700004445
700004454
700004613
700005083
WAT023486
WAT025531
Literature
References
Waters Global Services: W hy Choose Waters Services Brochure
Literature Reference 720001153EN
298
www.waters.com
Description
part no.
201000199
205000505
410000570
430002032
430002121
700001529
700001530
700004425
700004430
700004431
700004436
700004445
700004454
700004607
700004613
WAT023486
WAT025531
Description
Part No.
201000210
www.waters.com
205000505
405006414
405008440
430002032
430002121
700001529
700001530
700001836
700004425
700004430
700004431
700004439
700004445
700004607
700005084
WAT023486
WAT025531
WAT070122
299
Description
Part No.
WAT052675
700000253
WAT022946
WAT055631
WAT022628
WAT055632
WAT055633
WAT055629
WAT042200
Description
Part No.
WAT052673
WAT027629
WAT055631
WAT163-18
WAT022628
WAT055632
WAT055633
WAT055629
WAT042200
300
www.waters.com
Description
Part No.
WAT052671
WAS037497
WAT022615
WAT022626
WAT022628
WAT052119
WAT052220
WAT052230
WAT071406
WAT072919
Reagent Manager
Description
Part No.
201000102
700000146
700000145
www.waters.com
301
Description
Part No.
WAT052669
WAT045424
WAT077343
WAT073109
WAT077342
WAT078723
WAT078727
2707 Autosampler
Description
Part No.
700000680
700000683
700000684
700000685
700004063
700003872
700003921
700003922
700003923
700003924
700003925
700003928
700004086
700004088
700004089
700004090
Description
Part No.
201000196
302
www.waters.com
2747 Autosampler
Description
Part No.
201000132
Description
Part No.
205000105
205000106
205000107
205000108
205000109
205000112
205000115
205000116
205000117
205000118
205000119
205000134
700001397
www.waters.com
303
Description
Part No.
201000195
700000471
WAT272080
WAT272617
WAT272620
WAT272623
Description
Tabletop rack (for use with prep funnel rack)
Prep funnel (32 position, 2 each set)
Prep Funnel Rack (holds up to 4 prep funnels)
4-Microtiter Plate Rack
Multi-purpose rack
Carousel Rack, (2 x 2690 Carousels)
Eppendorf Tube Collection Rack
17mm OD Vial Collection Rack
28mm OD Vial Collection Rack
Standard Test Tube Rack, 120 positions
Prep funnel, 30 Position
Tygon Tubing, 6.35 mm ID x 9.52 mm OD x 5 m,
2/pkg (use with prep funnel rack)
Test Tube Rack
Teflon Tubing 8 mm ID x 50 ft (for use with prep funnel rack)
304
Part No.
289000440
725000106
725000107
725000110
725000113
725000144
725000145
725000146
725000147
725000152
WAT037046
WAT037047
WAT037072
WAT037090
www.waters.com
Manual Injectors
Description
Part No.
201000116
201000117
201000118
201000119
201000120
201000121
201000122
201000125
Description
Part No.
201000162
2777
700001230
700002210
700002292
700002293
700002297
700002298
700002439
Note: These rotors are not included in the PM Kit; must be ordered separately based
on the type of valve.
2777C
Literature
References
Waters Educational
Services Course Catalog,
Literature Reference 720000508EN
www.waters.com/courses
www.waters.com
305
Description
Part No.
WAS081142
WAS290423
WAS081346
WAT081159
WAT081321
WAT081158
WAT081157
289000614
700000923
700001786
205000321
205000322
205000323
Description
Part No.
201000186
306
Autopure flowcell
Dual path flowcell
I/O connector, 10 pin
Assembly, cable, ethernet, 10 ft straight through
Ethernet 10 ft crossover cable
MS flow cell rebuild kit
MS gasket kit, 10/pk
Analytical flowcell
Cuvette holder assembly
10 mm flowcell rebuild kit
3 mm flowcell rebuild kit
Gasket kit, 10/pk
10 mm cell, linearity solutions
Wavelength accuracy solutions
PQ test mix for absorbance detector
Two cuvette kit (empty)
Fuse, 3.25A, 250V 5 x 20 mm, fast-acting (5/pk)
Analog out cable assembly
Inert taper slit flow cell
Semi prep flowcell
Microbore flowcell
High pressure flowcell
Power Cord, 110V
289000614
289000935
323000247
441000372
700003423
700000168
700000169
WAS081140
WAS081333
WAS081346
WAS081347
WAS081348
WAT042881
WAT042885
WAT042887
700004155
700001800
WAT057235
WAT081157
WAT081158
WAT081159
WAT081321
WAT097194
www.waters.com
Description
Part No.
WAT052586
WAT057337
WAT057460
WAT057919
WAT057461
WAT057462
WAT057463
WAT057921
WAT057922
WAT057920
2996 Performance
Maintenance Kit
Description
Part No.
201000186
205000399
205000400
205000401
205000402
700001800
201000186
323000247
323000446
441000372
441000373
700003423
700001796
WAT057235
WAT021815
WAT021816
430001556
2998 Performance
Maintenance Kit
www.waters.com
307
maximum performance
Optimized electronic
features to ensure
reliable ignition and
longer lifetime
Independent certification
Part No.
WAS081142
WAT052586
201000131
201000186
201000159
T his offer cannot be combined with any other discounts or promotions. Not available to distributors or companies that resell spare parts (3rd party vendors).
Special promotion available only for the products listed above. Limited time offer.
Description
Part No.
700001183
700001710
WAT022384
WAT022385
WAT025604
WAT026805
WAT026973
WAT057235
WAT087141
WAT097194
WAT270868
Description
Part No.
201000159
415000415
700000574
Description
Flowcell Kit: 0.7 mm GC Working Electrode, Salt Bridge Reference Electrode
Flowcell Kit: 2 mm GC Working Electrode, Salt Bridge Reference Electrode
Flowcell Kit: 2 mm GC Working Electrode, ISAAC Reference Electrode
Flowcell Kit: 3 mm Pt Working Electrode, ISAAC Reference Electrode
Flowcell Kit: 3 mm AU Working Electrode, HyREF Reference Electrode
Flowcell Kit: 3 mm Ag Working Electrode, HyREF Reference Electrode
Fuse, 5 x 20, 2.5A, T 250V
RS-232 Cable
Dummy FlowCell
External I/O Cable
www.waters.com
Part No.
205004100
205004115
205004215
205004220
205004325
205004330
700001004
700001942
700001943
700001948
Part No.
700001949
700001950
700001951
700001952
700001953
700001954
700001955
700001956
700001957
700001958
700001959
700001960
700001961
700001962
700001963
700001964
700001965
700001968
700001985
700001994
700002003
700002069
700002103
309
Description
Part No.
201000131
700001840
700001618
Description
Part No.
WAT025604
WAT043069
WAT097194
WAT097332
WAT097410
Other Systems
Alliance GPC/V 2000 System
Description
Part No.
201000111
PM Kit includes: Plungers, Check valves, Seals, Filters, Needle, and Syringe (5 mL).
Parts & Accessories
Vial holder assembly, 10 mL (6/pk)
Filter vial assembly (6/pk)
Filter vial plunger assembly (6/pk)
GPC 2000 carousel handle assembly
GPC 2000 20 mm cap with septa (100/pk)
310
600000227
600000228
600000229
700000850
600000138
Carousel 24 position
700002406
www.waters.com
Part No.
Q-Tof Performance Maintenance Kit (for Q-Tof II, Q-Tof API US)
Q-Tof Ultima API Performance Maintenance Kit
Q-Tof Micro Performance Maintenance Kit
Q-Tof Premier Performance Maintenance Kit
Q-Tof Premier MALDI Performance Maintenance Kit
201000219
201000224
201000219
201000187
201000187
PM Kit Contains:
Source components
ESI probe components
Vacuum pump components
Part No.
201000187
PM Kit Contains:
Source components
ESI probe components
Vacuum pump components
Part No.
201000229
201000230
PM Kit Contains:
Source components
ESI probe components
Vacuum pump components
www.waters.com
311
Part No.
201000203
201000204
201000205
201000206
PM Kit Contains:
Source components
ESI probe components
Vacuum pump components
Part No.
201000238
201000239
201000240
201000241
PM Kit Contains:
Source components
ESI probe components
Vacuum pump components
312
www.waters.com
Part No.
176002055
PM Kit Contains:
Source components
ESI probe components
Vacuum pump components
Related Parts and Accessories not included in PM Kit:
APCI probe parts kit
IonSABRE spare parts kit
MUX 4/5 PM Kit
700000338
M956335DC2
201000149
Part No.
Quattro Ultima Pt
176002055
PM Kit Contains:
Source components
ESI probe components
Vacuum pump components
Related Parts and Accessories not included in PM Kit:
APCI probe parts kit
IonSABRE spare parts kit
MUX 4/5 Kit
700000338
M956335DC2
201000149
Part No.
176002050
PM Kit Contains:
Source components
ESI probe components
Vacuum pump components
Related Parts and Accessories not included in PM Kit:
MUX 4/5 parts kit
201000149
Part No.
201000170
PM Kit Contains:
Source components
Vacuum pump components
www.waters.com
313
Part No.
176002053
176002054
PM Kit Contains:
Source components
ESI probe components
Vacuum pump components
Related Parts and Accessories:
IonSABRE spare parts kit
MUX 4/5 parts kit
APCI probe parts kit
M956335DC2
201000149
700000338
TQ Detector
Description
Part No.
176002064
176002135
PM Kit Contains:
Source components
ESI probe components
Vacuum pump components
Part No.
314
176002049
www.waters.com
Xevo TQ
Description
Part No.
176002058
176002059
Related Parts
APCI Probe Service Kit
Outer Appi Source Service Kit
700004673
700004730
Xevo TQ-S
Description
Part No.
201000236
201000235
PM Kit Contains:
Source components
ESI probe components
Vacuum pump components
www.waters.com
315
Part No.
176002050
PM Kit Includes:
Source components
ESI probe components
Vacuum pump components
Related Parts and Accessories not included in PM Kit:
APCI probe spare parts kit
700000338
EMD 1000
ZQ Mass Spectrometer
Part No.
176002064
176002135
PM Kit Includes:
Source components
ESI probe components
Vacuum pump components
Part No.
176002064
176002135
PM Kit Includes:
Source components
ESI probe components
Vacuum pump components
Related Parts and Accessories not included in PM Kit:
SQD/TQD/3100 - ASAP Accessory
176002049
SQ Mass Detector
316
www.waters.com
Part No.
201000178
PM Kit Includes:
Source Components
Vacuum Pump Components
Part No.
176002062
176002063
PM Kit Includes:
Source components
ESI probe components
Vacuum pump components
Related Parts and Accessories not included in PM Kit:
APCI probe spare parts kit
IonSABRE spare parts kit
LockSpray source parts kit
MUX 4/5 parts kit
700000338
M956335DC2
201000144
201000149
Part No.
176002060
176002061
PM Kit Includes:
Source components
ESI probe components
Reference probe components
Vacuum pump components
Related Parts and Accessories not included in PM Kit:
APCI probe spare parts kit
IonSABRE spare parts kit
MUX 4/5 parts kit
700000338
M956335DC2
201000149
Literature
References
Edwards XDS Dry Vacuum Scroll Pump Conversion for
LCT Premier, LCT Premier XE Mass Spectrometers,
visit www.waters.com/upgrades
www.waters.com
317
Part No.
201000147
PM Kit Includes:
Source components
Vacuum pump components
Related Parts and Accessories not included in PM Kit
Field ionization/field desorption source PM Kit for GCT
201000146
Part No.
201000188
PM Kit Includes:
Source components
Vacuum pump components
Related Parts and Accessories not included in PM Kit
Field ionization/field desorption source PM Kit for GCT Premier
201000189
Part No.
201000145
PM Kit Includes:
Source components
Vacuum pump components
Related Parts and Accessories not included in PM Kit:
Electron impact (EI) source for Autospec
Chemical Ionization (CI) source for Autospec
Alternate CI/EI (ACE) source for Autospec
Field desorption (FD) source for Autospec
LSIMS/CS Gun source for Autospec
318
201000152
201000151
201000153
201000155
201000150
www.waters.com
[ bE aSSUREd ]
Choose Waters Global Services
Advance the Business of Science
with Empower Driven Services
Optimize laboratory resources through continuous improvement and
life cycle asset management. Empower Driven Services integrates
instrument services and the business analytics managed by Empower
Software to uncover efficiencies and cost savings. Learn more at
www.waters.com/empowerdriven.
Description
Part No.
205000343
700002775
Description
Part No.
700003776
700003775
700003779
700003780
700003777
700003778
T hese endfittings and accessories can be used with 4.6 mm, 4.0 mm, or
3.0 mm i.d. cartridges.
1
Description
Part No.
WAT037525
WAT023401
WAT037560
Column i.d.
Porosity
Part No.
1 mm
1 mm
2.1 mm
2.1 mm
3.0, 3.9, 4.6 mm
3.0, 3.9, 4.6 mm
7.8 mm
7.8 mm
19 mm
30 mm
2 m
0.5 m
2 m
0.5 m
2 m
0.5 m
2 m
5 m
2 m
2 m
600000175
600000176
600000177
600000178
600000179
600000180
600000181
600000182
600000183
600000184
320
Description
Qty.
Part No.
2/pk
5/pk
5/pk
2/pk
2/pk
1/pk
1/pk
PSS614100
PSS614103
PSS614104
PSS614102
PSS614101
PSS614108
WAT015674
10/pk
PSS614113
PSS830008
www.waters.com
SLIPFREE Connectors
New Generation HPLC Column Connector
SLIPFREE Fittings
Part No.
PSL618000
PSL618004
PSL618006
PSL618008
PSL618010
PSL618001
PSL618003
PSL618005
PSL618007
PSL618009
PSL618011
Single SLIPFREE
Double SLIPFREE
Guard Column
or Filter
HPLC Column
Injector
Detector
www.waters.com
321
0.500 in.
0.826 in.
PSL613315
Part No.
PSL613315
0.375 in.
0.698 in.
PSL613322
0.125 in.
0.235 in.
PSL613316
Part No.
PSL613322
PSL613316
0.125 in.
0.275 in.
PSL613302
1/4 in.
0.605 in.
We offer both fingertight and hex-head nuts for use with Double-Ferrules.
T he fingertight version may be hand-tightened for operation at up to
6,000 psi. Use the hex-head version for connections that are difficult to
reach or closely spaced.
T hese fittings fit virtually any female 1/16 in. fittingParker, Swagelok,
Waters, Valco, Rheodyne, Upchurch, etcall having 10-32 threads.
PSL613324
0.475 in.
0.700 in.
PSL613301
322
Part No.
PSL613302
PSL613324
PSL613301
PSL613325
1/2 in.
0.525 in.
PSL613325
www.waters.com
Length/Material
Part No.
0.125 (3.2)
0.149 (3.8)
0.250 (6.3)
0.080 (2.0)
0.178 (4.52)
0.149 (3.8)
0.125 (3.2)
0.125 (3.2)
0.125 (3.2)
0.062 (1.57)
0.062 (1.57)
0.062 (1.57)
0.119 (30.0)
0.190 (4.8)
0.058 (1.5)
0.148 (3.76)
0.119 (30.0)
0.020 (0.508)
0.009 (0.228)
0.040 (1.0)
0.009 (0.228)
0.040 (1.0)
WAT026808
WAT026809
WAT026810
WAT026974
WAT051041
WAT051052
WAT088430
WAT088431
WAT088432
WAT088561
WAT088563
WAT005137
Description
Part No.
WAT005063
WAT005070
WAT005079
WAT007020
WAT022330
WAT021817
WAT025313
WAT025566
WAT025604
WAT046-12
WAT021812
WAT021815
WAT060051
WAT021816
WAT075215
WAT084078
WAT097332
Length/Material
Part No.
0.0625 (1.6)
0.0625 (1.6)
0.0625 (1.6)
0.0625 (1.6)
0.125 (3.2)
0.125 (3.2)
0.0625 (1.6)
0.005 (0.127)
0.020 (0.508)
0.030 (0.762)
0.040 (1.020)
0.062 (1.57)
0.093 (2.36)
0.009 (0.228)
10 ft. (3 m), SS
10 ft. (3 m), SS
10 ft. (3 m), SS
10 ft. (3 m), SS
10 ft. (3 m), SS
10 ft. (3 m), SS
10 ft. (3 m), SS
WAT241039
WAT026804
430000366
WAT026805
WAT026806
WAT026807
WAT026973
Length/Material
Part No.
0.0625 (1.6)
0.0625 (1.6)
0.0625 (1.6)
0.0625 (1.6)
0.005 (0.127)
0.010 (0.254)
0.015 (0.381)
0.020 (0.508)
WAT022995
WAT022996
WAT022997
WAT022998
WAT031795
WAT022999
WAT026-04
WAT022384
WAT022385
Filters
Last Drop Mobile Phase Filters
T he Last Drop Mobile Phase Filter utilizes a flat filter element whic h
sits parallel to the bottom of the reservoir. T his design allows the filter
to draw all but the last 2% of the mobile phase from the reservoir
without drawing air into the system. Last Drop Filters are available with
316L Stainless Steel or PAT (PEEK alloyed with Teflon) filter elements
in inert Teflon housings. T he top of the housing has a PEEK tripod which
fits into 1.5, 2.2 or 3.5 mm id pump inlet lines.
www.waters.com
Part No.
PSL901290
PSL901291
PSL901292
PSL901294
Universal PEEK
Tubing Adapter
Stainless Steel
or PAT
Filter Element
323
10-32 T hread
Description
Part No.
PEEK Union with 2 PEEK Fingertight Nuts and Double Ferrules 1/16 in.
PEEK Union without Nuts and Ferrules 1/16 in.
PEEK TEE with One Piece Fingertight Fitting
PEEK CROSS with One Piece Fingertight Fitting
PEEK TEE Without Fittings
PEEK CROSS Without Fittings
PEEK One Piece Fingertight Fitting
PSL613312
PSL613313
PSL613317
PSL613319
PSL613318
PSL613320
PSL613315
PEEK union
Part No.
PSL613321
Qty.
Part No.
1/pk
10/pk
PSL618021
PSL618022
324
www.waters.com
Description
Part No.
PSL901282
Description
Solvent Inlet Filter Kit - Contains one 3 in. length of Teflon Tubing,
1 Plastictight Fitting, 5 Replacement 10 m Filters
Plastictight Fitting with Teflon Tubing
1/16 in. i.d. x 1/8 in. o.d. x 3 ft.
Replacement Filter 10 m 5/pk
Part No.
PSL613570
PSL613578
Solvent Inlet Filter 10 m with 1/16 in. o.d. stem for 1/8 in. o.d. tubing
Solvent Inlet Filter 10 m with flangeless fittings for 1/8 in. o.d. tubing
Part No.
PSL613609
PSL613607
PSL613608
PSL613609
www.waters.com
Stainless
Steel Stem
Part No.
PSL613457
325
326
www.waters.com
[ Indices ]
Alphabetical Index....................................................................................................................................................................................328
Part Number Index ....................................................................................................................................................................................336
327
aLPhabETICaL INdEx
ACQUITY UPLC
ACQUITY CSH Columns ..................................................................................... 91
Guard-Pak Holder..............................................................................................165
Resolve Columns...............................................................................................161
SAM Reagents...................................................................................................170
ELS Detector......................................................................................................285
Sample Manager...............................................................................................285
Analytical Chromatography
328
Alphabetical Index
www.waters.com
aLPhabETICaL INdEx
Waters Spherisorb Columns .............................................................................152
XSelect Columns...............................................................................................118
XTerra Columns.................................................................................................140
Atlantis
Delta-Pak
Bulk Packings....................................................................................................192
HSS T3 ..............................................................................................................135
Introduction.......................................................................................................132
Preparative Columns.........................................................................................159
T3 Bonding .......................................................................................................133
LC and MS Standards........................................................................................255
2475 Multi-Wavelength
Fluorescence Detector ......................................................................................310
Cation-exchange Columns................................................................................213
DNA/RNA/Oligonucleotides .............................................................................246-248
Bondapak/Bondapak
Bulk Packings....................................................................................................191
Preparative Columns.........................................................................................177
Extraction Manifold............................................................................................. 38
F
Fast Fruit Juice Analysis Column...............................................................................173
Capillary Columns
and Carbohydrates......................................................................................................174
Atlantis .............................................................................................................260
Ferrules .......................................................................................................................322
Symmetry300 ..................................................................................................260
Symmetry .........................................................................................................260
Filtration.................................................................................................................. 80-84
XTerra ................................................................................................................260
www.waters.com
Alphabetical Index
329
aLPhabETICaL INdEx
Pall Life Sciences Part Number Cross Reference ............................................... 83
Pall Life Sciences Solvent Filtration Product List ............................................... 83
Pall Life Sciences Syringe Filters Product List ................................................... 84
Sirocco Protein Precipitation Filtration Plate..................................................... 80
H
Handilok CTFE Fittings ...............................................................................................324
Heat Sealer for 96-Well Collection Plates .................................................................. 67
High Performance Carbohydrate Cartridge Column .................................................170
HILIC ...........................................................................................................................136
IC-Pak
Introduction.......................................................................................................268
L
Lanthanide Analysis ...................................................................................................175
Last Drop Mobile Phase Filters ...................................................................................323
Manifolds
Guard Products
Waters Spherisorb Guard Columns,
Guard-Pak Holder..............................................................................................165
Guard-Pak Inserts ..............................................................................................165
Guide to Guard Column Selection and Use .....................................................163
Sentry Guard Columns Product List ..................................................................164
Sentry Guard Holders Product List ...................................................................163
330
Alphabetical Index
MassPREP
MassPREP Consumables for MS Analysis ..............................................253-256
MassPREP Digestion Standard Mixtures..........................................................256
MassPREP Glycoanalysis Kit............................................................................252
MassPREP MS and LC/MS
Consumables for Biomolecules ........................................................................253
www.waters.com
aLPhabETICaL INdEx
MassPREP On-Line Desalting Cartridges.........................................................258
NanoEase
Mass Spectrometers
Oasis
Elution Plates.................................................................................................... 26
nanoACQUITY UPLC
PEEK
www.waters.com
Alphabetical Index
331
aLPhabETICaL INdEx
PEEK Unions, Tees, and Crosses.......................................................................324
PrepLC
Protein-Pak
HIC Columns......................................................................................................237
PTFE/ETFE Tubing and Fittings ..................................................................................323
Pumps
Porasil/Porasil
Preparative Columns.........................................................................................191
QuEChERS ..................................................................................................................... 54
Introduction.......................................................................................................177
RapiGest SF .................................................................................................................254
Resolve
Sample Preparation
332
Alphabetical Index
www.waters.com
aLPhabETICaL INdEx
Sample Vials and Accessories
Vial Descriptions................................................................................................. 78
Styragel
HR High-Resolution Columns...........................................................................272
Sentry
Sep-Pak
96-Well Plates.................................................................................................... 42
Cartridge Accessories......................................................................................... 51
Cartridge Connections Kit .................................................................................. 51
Cartridge Selection Guide .................................................................................. 42
Certified Sep-Pak Solid-Phase Extraction Cartridges .................................. 48-49
Chemistry Selection Chart............................................................................ 42-43
Chemistry Selection Guide ................................................................................. 40
DNPH Silica Cartridges ....................................................................................... 45
Dry SPE Cartridges ............................................................................................. 47
Ozone Scrubber Cartridge.................................................................................. 46
Specialty Chemistries ......................................................................................... 44
Vac RC SPE Cartridges Product List .............................................................. 38-39
Vac SPE Cartridges Product List ................................................................... 38-39
Waters Positive Pressure-96 Processor ...................................................... 38, 51
X PoSure Aldehyde Sampler Cartridges............................................................. 46
Separations Modules
Preparative Columns.........................................................................................190
www.waters.com
SymmetryPrep
Columns for Purification and Isolation ............................................................190
SymmetryShield
Introduction.......................................................................................................147
Preparative Columns.........................................................................................190
Syringe Filters............................................................................................................... 84
Alphabetical Index
333
aLPhabETICaL INdEx
T3 Columns .................................................................................................................133
Trapping Columns
Atlantis .............................................................................................................259
Symmetry .........................................................................................................259
Vial Descriptions................................................................................................. 78
Symmetry300 ..................................................................................................259
SCX ....................................................................................................................259
Ultrahydrogel
Aqueous SEC Columns .....................................................................................279
Column Selection Guide ...................................................................................270
For Sugar and Carbohydrate Analysis .............................................................167
Ultrastyragel
For Cholesterol Analysis ..................................................................................173
Preparative GPC Columns .................................................................................276
UPLC
ACQUITY CSH Columns ..................................................................................... 91
ACQUITY UPLC BEH HILIC Columns .................................................................. 93
ACQUITY UPLC BEH Reversed-Phase Columns ................................................ 93
ACQUITY UPLC Column Ordering Information ................................................. 99
ACQUITY UPLC HSS Columns............................................................................ 95
Heat Seal for ACQUITY UPLC Systems ....................................................67, 105
Introduction......................................................................................................... 90
nanoACQUITY UPLC 2D Kit .............................................................................262
nanoACQUITY UPLC Column Ordering Information.......................................262
nanoACQUITY UPLC Columns .........................................................................261
Plates for ACQUITY UPLC Systems ..........................................................66, 104
Screw Cap 12 x 32 mm Vials for ACQUITY UPLC Systems ...................64, 103
Settings for ACQUITY UPLC Vials ..................................................................... 65
Vanguard Pre-Columns ..............................................................................96, 102
Waters ACQUITY UPLC System Sample Manager ...................................65, 104
USP L Column Listing .............................................................................................113
W
Waters Global Services...............................................................................................319
Waters Positive Pressure-96 Processor ................................................................ 38, 51
Waters Quality Parts and Supplies
Introduction.......................................................................................................284
ACQUITY UPLC System PM Kits
2996 PDA Detector ...............................................................................285
ACQUITY I2V BSM PM Kit ......................................................................285
Binary Solvent Manager .........................................................................285
ELS Detector............................................................................................285
FLR Detector ...........................................................................................285
Sample Manager.....................................................................................285
Spare Parts & Tubings ............................................................................285
TUV Detector ..........................................................................................285
TUV/PDA/eLambda Detector..................................................................285
ACQUITY UPLC Local Console Controller Kit ..................................................289
nanoACQUITY UPLC System
Binary & Auxiliary Solvent Manager PM Kit .........................................290
High Pressure BSM/ASM Spares & Tubings ...........................................291
High Pressure SM Spares & Tubings ......................................................291
nanoACQUITY UPLC 2D Spares & Tubing.............................................291
nanoACQUITY UPLC Columns ...............................................................261
Sample Manager PM Kit .........................................................................290
TUV Detector PM Kit ...............................................................................290
TUV Detector Spares & Tubings.............................................................291
HPLC Pumps
334
Alphabetical Index
www.waters.com
aLPhabETICaL INdEx
1525 HPLC Pump ..................................................................................297
Gradient Modules
XBridge
Analytical Columns ..........................................................................................127
Introduction.......................................................................................................122
717plus Autosampler............................................................................302
Detectors
2414 Refractive Index Detector ............................................................309
XSelect
Analytical Columns ..........................................................................................118
Introduction.......................................................................................................118
www.waters.com
Alphabetical Index
335
176001519.............................. 258
176001882.............................. 101
176002140................................ 99
186000150.............................. 146
176000862................................ 99
176001883.............................. 101
176002141................................ 99
186000151.............................. 146
176000863................................ 99
176001884.............................. 101
176002142................................ 99
186000152.............................. 146
176000864................................ 99
176001885.............................. 101
176002143................................ 99
186000153.............................. 146
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176002165................................ 99
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186000192.............................. 150
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186000194.............................. 150
176001092.............................. 100
176001802................................ 99
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186000100.............................. 148
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176001093.............................. 100
176001803................................ 99
186000101.............................. 148
176001094.............................. 100
176001804................................ 99
176002050............................ , 313
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176001805................................ 99
176002053.............................. 314
186000103.............................. 148
186000199.............................. 150
176001806................................ 99
176002054.............................. 314
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176002055.............................. 313
186000105.............................. 148
176001808................................ 99
176002058.............................. 315
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176001809................................ 99
176002059.............................. 315
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176001810.............................. 100
176002060.............................. 317
186000108.............................. 148
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176001811.............................. 100
176002061.............................. 317
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176001812.............................. 100
176002062.............................. 317
186000110.............................. 148
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176001813.............................. 100
176002063.............................. 317
186000111.............................. 148
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186000115.......................... 35, 36
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176002126.............................. 101
186000116.......................... 35, 36
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176001304................................ 99
176002127.............................. 101
186000117.......................... 35, 36
186000217.............................. 148
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176002128.............................. 101
186000118.......................... 35, 36
186000218.............................. 148
176001310................................ 99
176001821.............................. 100
176002129.............................. 101
186000128.......................... 34, 36
186000223.............................. 148
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186000132.......................... 35, 36
186000224.............................. 148
176001367.............................. 262
186000145.............................. 148
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176002136................................ 99
186000146.............................. 148
186000234................................ 71
176001398.............................. 100
176002137................................ 99
186000147.............................. 148
186000235.............................. 190
176001459.............................. 243
176002138................................ 99
186000148.............................. 146
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176002139................................ 99
186000149.............................. 146
186000237.............................. 190
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186000420.............................. 142
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186000535.............................. 142
186000366.......................... 34, 37
186000480.............................. 142
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186000481.............................. 142
186000536.............................. 142
186000425.............................. 142
186000482.............................. 142
186000537.............................. 142
186000426.............................. 142
186000483.............................. 142
186000538.............................. 141
186000427.............................. 142
186000484.............................. 142
186000539.............................. 141
186000428.............................. 142
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186000540.............................. 141
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186000430.............................. 142
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186000542.............................. 141
186000431.............................. 142
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186000543.............................. 141
186000432.............................. 142
186000489.............................. 142
186000544.............................. 141
186000433.............................. 142
186000490.............................. 142
186000545.............................. 141
186000434.............................. 142
186000491.............................. 142
186000546.............................. 141
186000435.............................. 142
186000492.............................. 142
186000547.............................. 141
186000436.............................. 142
186000493.............................. 142
186000548.............................. 141
186000437.............................. 142
186000494.............................. 142
186000549.............................. 141
186000438.............................. 142
186000495.............................. 142
186000550.............................. 141
186000439.............................. 142
186000496.............................. 142
186000551.............................. 141
186000440.............................. 142
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186000441.............................. 142
186000498.............................. 141
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186000442.............................. 142
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186000443.............................. 142
186000500.............................. 141
186000555.............................. 142
186000446.............................. 141
186000501.............................. 141
186000556.............................. 142
186000447.............................. 141
186000502.............................. 141
186000557.............................. 142
186000448.............................. 141
186000503.............................. 141
186000558.............................. 142
186000449.............................. 141
186000504.............................. 141
186000559.............................. 142
186000450.............................. 141
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186000508.............................. 141
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186000454.............................. 141
186000509.............................. 141
186000564.............................. 142
186000455.............................. 141
186000510.............................. 142
186000565.............................. 142
186000456.............................. 141
186000511.............................. 142
186000566.............................. 142
186000457.............................. 141
186000512.............................. 142
186000567.............................. 142
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186000513.............................. 142
186000568.............................. 142
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186000514.............................. 142
186000569.............................. 142
186000460.............................. 141
186000515.............................. 142
186000570.............................. 142
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186000238.............................. 190
186000252.......................... 34, 36
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186000259.......................... 34, 36
186000261.......................... 35, 36
186000262........................ , 36, 38
186000268.............................. 146
186000269.............................. 146
186000270.............................. 146
186000271.............................. 146
186000272C .................. 70, 73, 74
186000272DV ...................... 70, 74
186000273.......................... 70, 74
186000273DV ...................... 70, 74
186000274.......................... 70, 74
186000275...................... 150, 164
186000276.............. 150, 214, 235
186000277.............. 150, 214, 235
186000278.............. 150, 214, 235
186000279.............. 150, 214, 235
186000367.......................... 34, 37
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186000373.......................... 34, 37
186000375.......................... 34, 37
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186000380.......................... 35, 36
186000381.......................... 35, 36
186000382.......................... 35, 36
186000383.......................... 34, 36
186000384C .............. 240, 243, 70
186000384DV ............................ 70
186000385C ........................ 68, 70
186000385DV ...................... 68, 70
186000386.............................. 141
186000387.............................. 141
186000388.............................. 141
186000389.............................. 141
186000390.............................. 141
186000391.............................. 141
186000392.............................. 141
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186000572.............................. 142
186000302................................ 71
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www.waters.com
337
186000695.............................. 146
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186000711.............................. 190
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186000776.......................... 35, 36
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186000832.............................. 143
186000833.............................. 143
186000834.............................. 143
186000835.............................. 143
186000836.............................. 143
186000837C .............................. 72
186000837DV ............................ 72
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186000838DV ...................... 72, 76
186001126DV ............................ 74
186001181.............................. 141
186001127................................ 75
186001183.............................. 142
186001128C .............................. 74
186001184.............................. 142
186001128DV ............................ 74
186001185.............................. 141
186001130C .............................. 74
186001186.............................. 141
186001130DV ............................ 74
186001187.............................. 141
186001131C .............................. 74
186001188.............................. 141
186001131DV ............................ 74
186001189.............................. 141
186001190.............................. 142
186001191.............................. 142
186001192.............................. 141
186001193.............................. 141
186001135.......................... 72, 76
186001194.............................. 142
186001135DV ............................ 76
186001195.............................. 142
186001139.............................. 142
186001196................................ 36
186001140.............................. 142
186001203.............................. 142
186001141.............................. 142
186001204.............................. 142
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186001023.............................. 189
186001158.............................. 189
186001291.............................. 137
186000900.............................. 141
186000925.............................. 158
186000926...................... 213, 234
186000965.......................... 72, 76
186000979.............................. 141
186000982.............................. 143
186000984.......................... 71, 75
186000984DV ...................... 71, 75
186000988................................ 36
186000989................................ 70
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186000999.......................... 72, 76
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186001004.............................. 189
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186000840.......................... 72, 76
186001025.............................. 189
186001160.............................. 189
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186000679.......................... 34, 36
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186001026.............................. 189
186001161.............................. 189
186001297.............................. 137
186000683.......................... 35, 36
186000841.......................... 72, 76
186001027.............................. 189
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186001164.............................. 189
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186000694.............................. 148
338
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186001480.............................. 189
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186001831........................ 209, 38
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186001346.............................. 137
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186002035................................ 36
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186001879.......................... 34, 36
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186002036................................ 36
186001371.............................. 188
186001761.............................. 275
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186001771.............................. 275
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186002047................................ 36
186001393.............................. 137
186001772.............................. 275
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186002048................................ 36
186001395.............................. 137
186001773.............................. 275
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186002049................................ 36
186001413................................ 36
186001774.............................. 275
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186002051................................ 36
186001414................................ 36
186001775.............................. 275
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186002052................................ 37
186001420.......................... 73, 76
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186002053................................ 37
186001421................................ 76
186001777.............................. 275
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186002054................................ 37
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WAT200841 ................................ 84
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WAT200846 ................................ 84
WAT201549 .............................. 213
WAT210579 .............................. 148
WAT210582 .............................. 148
WAT210585 .............................. 148
WAT210588 .............................. 148
WAT210591 .............................. 148
WAT210594 .............................. 148
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WAT241039 .............................. 323
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Dear Colleague,
Our catalog contains todays leading separations technology products. With over 50 years of market
leadership we remain committed to the development and manufacture of products that solve todays most
challenging separations problems to enable our customers success. We are driven by our core mission to
provide enabling technologies in products that set the industry standard for performance and reproducibility.
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experience has been improved to make it easier and faster for you to obtain pricing and availability when
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T his year for the fist time our catalog is available in digital form. You may access and download it at
www.waters.com/catalog.
T his past year we launched several innovative, enabling products including our new CSH technology
that is the basis for the XSelect brand of HPLC columns and the ACQUITY CSH brand of UPLC columns,
the Ostro Sample Preparation plate for the removal of phospholipids in biological samples and the
Viridis line of columns for supercritical fluid chromatography.
To learn more about how we can help you achieve your objectives please telephone one of our 54 worldwide offices and interact with our highly trained account specialists.
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Taiwan
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All Other Countries
Waters Corporation
34 Maple Street
Milford, MA 01757 USA
Tel: 508 478 2000
800 252 4752
Fax: 508 872 1990
Michael J. Yelle
Senior Director, Chemistry Commercial Operations
Waters Corporation Milford, MA, U.S.A.
www.waters.com
720003820EN
2011 / 2012
www.waters.com
2011/2012