To separate solutes differing in vapor pressure and/or intensity of solute-stationary phase interactions

What are main components of a gas chromatograph ?

Gas chromatograph

Mobile phase
Carrier gas: N2, He, H2 non-solvating, high purity, and chemically inertness

Thin film

Thick film

Pure He, N2 (> 99.995%)

Further purifying carrier gas by humidity, hydrocarbon, and oxygen traps/filters

Gas-Liquid Chromatogr. (GLC)

Stationary phase: immobilized liquid

GC
Gas-Solid Chromatogr. (GSC)
Stationary phase: solid

GC stationary phase (GSC)

Stationary phase: Al2O3, carbon Less popular than GLC Disadvantages : long retention times, tailing, poor reproducibility from col. to col. Main applications: gas analysis

GC stationary phase (GLC)

Features of good liquid stationary phases for GLC unreactive, high thermal stability low vapor pressure good coating characteristics wide temperature operating range

What are stationary phases of GC ?

Stationary phase (GLC)

High molecular weight hydrocarbons - Squalane (C30H62) (shark liver oil )
Proposed substitution of apolane-87 for squalane as a nonpolar reference phase in gas chromatography Analytica Chimica Acta Vol. 225, 1989, 193-203 Squalane is the standard, nonpolar reference phase . SP in GLC ... Primary objections to its use as a reference phase are its high volatility, compositional variations due to the presence of impurities, susceptibility to oxidative degradation, poor film forming. the synthetic hydrocarbon Apolane-87, its use as a reference phase is to be preferred.

- Apolane-87 (C87H176)

Stationary phase (GLC)

Poly (siloxane)

C BH

poly(siloxane) polymer

poly(silarylene-siloxane) copolymer

poly(carborane-siloxane) copolymer

Stationary phases
Stationary phases can be deposited on to column walls by cohesive or wetting forces surface-bonded (bonded phase) it may be cross-linked to produce a liner or envelope of stationary phase polymer within the column it may be both cross-linked and surface-bonded
Cross-linking can produce a stationary phase that is non-extractable (splitless, on-cloumn injection or washing the columns)

Polarity

Effect of stationary phase polarity on separation of (i) An alkane and an ester of similar volatility (68 oC) (ii) An alkane and 2 esters of different volatility (68 oC and 57oC)

Packed and capillary column

Packed column

Capillary column

GC columns
Packed columns
1-3mm i.d. < 5m length 0.1-0.4 mm dp

Capillary columns
0.1-0.33 mm i.d. (mega-bore 0.53 mm) 12-150 m length 0.1 -0.5 m film thickness

Wall Coated Open Tubular column (WCOT)

Porous Layer Open Tubular column (PLOT)

Support Coated Open Tubular column (SCOT)

Wall Coated Open Tubular column (WCOT)

Fused silica tube 0.10.5 mm i.d.

Chemically bonded phase

0.1 - 5 m

Typical characteristics of GC columns

Preparation of capillary column

Fused silica

Coater 2

Curing oven 2 Furnace

(2300oC)
Coater 3

Curing oven 3

10m tower
Coater 1

Coater 4

(Polyimide)
Curing oven 4

Curing oven 1

Reel

Purity of silica and its strength

clean room conditions are required for preparation of fused silica !

Modified silica

Fused silica

thin-walled columns (0.20 mm ID x 0.25 mm OD) that were inherently straight, extremely strong, and highly flexible are made from fused silica.

Column deactivation
before after
C10 C10 C12 2 C12 3 1 4 6

C17

C17

1 3,5-dimethylpyrimidine 2 C8-NH2 3 2,6-dimethylaniline

4 C10-NH2 5 N,N- dicyclohexylamine 6 C12-NH2

Column thermal stability

g/min

Bleeding as a function of temperature

Temperature

Standard bleed test

Thermally-induced catalytic stationary phase decomposition due to alkali metal ions

Scotch Whisky *

Packed Col. 5% Carbowax 20M on 80/120 Carbopack B (2 m 2 mm I.D.) FID; Temp. : 70 C, 4C/min. to 150 C

Scotch Whisky production

Malting mashing fermentation distillation maturation

Barley

Malted barley

Distilled lime oil

-Fenchyl alcohol, -Phellandrene, -Pinene, -Bisabolene, -Caryophyllene, -Terpinene, 1,4-Cineole, d-Limonene, p-Cymene, trans--Bergamotene, Borneol, Camphene, Decanal, Dodecanal, Geranial, Geranyl acetate, Linalool, Myrcene, Neral, Neryl acetate , Terpinen-1-ol, Terpinen-4-ol, Terpinolene, -Terpineol, natural, Kosher

Columns with small id. and thin film for FAST GC

Semivolatile analytes

Column: SLB-5ms, 20 m x 0.18 mm i.d., 0.18 m film thickness Oven: 40 C (0.7 min.), 55 C/min. to 240 C, 28 C/min. to 330 C (2 min.) Detector: MS

Sample introduction
(capillary column)
COMPLETE & FAST TRANSFER !!!! for sharp peaks - Injector volume - Injector temp. (instant vaporization) ) - Residence time (in split and splitless) - Deactivation of glass surface - Split ratio - Solvent Effect in splitless

On-column injection

Analytes: - Thermally labile - Low volatility

A large volume injection with RETENTION GAP

Wide bore col. as Retention gap

Flooded zone formed in the retention gap

Solvent evaporating from the rear of the flooded zone

Volatile components concentrated by solvent effect

Stationary phase

Less volatile components focused by stationary phase

Programmed-Temperature Vaporization (PTV)

Controlled and programmable injector temperature cooling (cool air, liquid N2, ) heating (hot air, , ~15 oC/s) Most solvent is removed through split valve at low temperature. Analytes will be introduced into the column when split valve is closed and the injector is rapidly heated

injection of large sample volumes (tens - 200 L )

Can be used as normal split/spitless, vapor sample introduction

Discrimination: low boiling point compounds > high boiling point compounds (n-alkanes -> up to C16)

Injection techniques for volatile compounds?

Static headspace
(for volatile analytes)
avoids lengthy and costly sample preparation step !

Static headspace
Partition Coefficient Phase Ratio K = Cs/Cg = Vg/Vs

Cg=Co/(K+)
volatile analytes

Decrease K by - Increasing temperature KEtOH(air/water): 1355 (40oC) ; 328 (80oC) - High inorganic salt concentration in aq. phase decrease solubility organic volatile analytes

Sample, dilution solvent & matrix modifier

Static headspace analysis of saturated short chain aldehydes from cardboard

(derived from lipid degradation) MS detection

1.5 g sample

Solid Phase Micro Extraction SPME

solventless sample preparation

For liquid and headspace.

Which injection modes should I select for my samples?

Split

Concentration Volatility Thermal stability Sample cleanness

Splitless On-column Purge and trap Headspace PTV split

Detectors
High sensitivity Fast response

Requirements for ideal detectors?

Wide linear dynamic range Universal/selectivity Stability Robustness Ease of operation

Thermal conductivity detector (TCD)

Universal (non-specific) detector Thermal conductivity of organic compounds are similar and very different from Helium Less sensitive

Flame Ionization Detector (FID)

Most popular Universal High sensitive (10-13 g C/sec) Wide linearity range (106-107) Robust

Contributions of structure to the response of FID

Electron Capture Detector (ECD)

Structure selective (halogen, nitro, nitrile)

63Ni,

emitter

Gas flow

N2 A + A-

N 2+ + A
N2+
-

base current decrease current

N2

ECD detector
Simple and reliable Sensitive to electronegative groups (halogens, nitro, nitrile) Largely non-destructive Limited dynamic range (102 - 103)

Relative response of the ECD to various organic compounds

Compounds 1-chlorobutane 1,4-dichlorobutane 1-bromobutane 1-iodobutane Chloroform Carbon tetrachloride Relative response 1.0 15.0 2.8 102 9.0 104 6.0 104 4.0 105

MS DECTECTOR
Electron Impact Ionization (EI)
-lactam

+ M M+ + 2 A+ B+ C+

MS library (70 eV) for identification of compounds

MS DECTECTOR
Base peak

Fragment ions

Molecular ion

Scan mode SIM (selected ion monitor)

qualitative quantitative

How to analyze highly polar compounds by GC ?

What is GC derivatization?
The process of chemically modifying a compound to produce a new compound which has properties that are suitable for analysis using a GC

Why derivatize?
Improve volatility, thermal stability, i.e. convert polar
compounds (acids, alcohols) to esters for higher volatilities so that they can be eluted at reasonable temperatures without thermal decomposition or molecular re-arrangement

Enhance detector response, i.e. tagging with halogen

for ECD detection

Main types of derivatization

Alkylation Silylation
H+ acid + alcohol ester + H2OH2O acid + alcohol ester +

Acylation

RCOCO-R + R'OH RCOOR' + R-CO-O-H

2,4-D in RICE MRL : 0.1mg/kg (WHO)

2,4-Dichlorophenoxyacetic acid BP: 160 oC

Esterification (methylation)
MeOH H2SO4

LOD: 3 ppb LOQ: 10 ppb

Multidimensional GC

(1) acetone, (2) 2-butanone, (3) benzene, (4) isopropylmethylketone, (5) isopropanol, (6) ethanol, (7) toluene, (8) propionitrile, (9) acetonitrile, (10) isobutanol, (11) 1-propanol, (12) butanol

Improve separation of complex mixtures (petroleum products, PCBs, enantiomers in flavor and food technology)

Multidimensional GC
Sections of a one-dimensional chromatogram will transferred and independently separated on a second column of different characteristics
Injector Modulator Detector Detector

Refocusing on the 2D column by modulators is crucial !!!

Heartcut GC x GC
C1: Poly(ethyleneglycol)

A B Cut
C2: Poly(dimethylsilosane)
C5 C6 C7

(1) acetone, (2) 2-butanone, (3) benzene, (4) isopropylmethylketone, (5) isopropanol, (6) ethanol, (7) toluene, (8) propionitrile, (9) acetonitrile, (10) isobutanol, (11) 1-propanol, (12) butanol

Comprehensive two-dimensional GC
TOTAL TRANSFER of all sample components from the first column to the second column as a series of pulses which are separated sequentially and individually on the second column 1D long (conventional) column 2D short column (for very fast separation, few seconds / run)

Cryogenic trap to refocus heart-cuts

1D separation

2D separation

2D GC separation

Illustration of how two overlapping peaks emerging from D1 (A) are resolved in GC x GC after passage to D2 (B).

Sources of activity of glass surface

Metal ions, boron Silanol groups
Vicinal silanol Si Free silanol Geminal silanol Si H Weak or none O O H

Si

OH
> 3.1m

Si

OH OH Hydrated silanol H O H

Si

OH Weak or none Si Si O O H H

Strong

Strong