28/04/2019 Classification of HPLC Separations by Column Dimensions

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POSTED ON NOVEMBER 26, 2013 BY DR. DEEPAK BHANOT 2 COMMENTS

Classi cation of HPLC Separations

by Column Dimensions
The classi cation of HPLC methods is governed by several factors such as :

Purpose of separation – identi cation, quanti cation or collection of puri ed fractions

Available sample size
Detection e ciency and choice of detector

Based on the available information on the sample and purpose of analysis HPLC methods are
classi ed into the following four basic modes :

Microbore HPLC
Analytical HPLC
Semipreparative HPLC
Preparative HPLC

Broadly speaking the classi cation is based on the following parameters :

Classi cation  id                       Length Packing Flow rate POPULAR LATEST

size                    
TODAY WEEK MONTH ALL
Millibore 1-2mm 15-30mm 1-3 μm 1-100μl/min
Analytical 4-0-4.6mm 150-250mm 3-5 μm 0-1-10 ml/min Jetpack plugin with Stats module
needs to be enabled.
Semi preparative 10-25 mm 150-250mm 10-20 μm 1-10ml/min
Preparative 25-75mm 150-250mm 20-50 μm 10-200ml/min

Microbore HPLC
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Majority of academic and industrial applications involve use of analytical HPLC columns. In such
situations sample is available in su cient quantity (pg-ng levels) to meet the analysis Spectroscopy (80)
requirements. Objective of analytical separations is qualitative identi cation followed by
Stability studies (8)
quantitative estimations. Common HPLC detectors such as UV, RI, Conductivity,
Electrochemical,etc provide satisfactory results in analytical separations. Training (21)

Weighing (3)

Semi Preparative HPLC

Semi Preparative HPLC Column

As you move from analytical to semi preparative mode sample quantities range between μg-mg
levels. The id of the column also gets larger. The technique is used for mainly separation and
fractional collection of component of pure sample components..

Preparative HPLC

Preparative HPLC Columns

Preparative separations are aimed at isolation and laboratory scale preparation of components
of the sample. Isolation can be done in mg-gm levels. The columns are much more broader.
Operational cost is signi cantly higher than other conventional separation methods so it is used
for isolation of rare or expensive products such as ultrapure standards, trace impurities,
metabolites and biological materials such as peptides

In the coming time advancements in column packings and instrumentation will lead to more
e cient and time-saving identi cation, quanti cation and separations but it seems unlikely that
there will be further changes in the column dimension based classi cation.

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28/04/2019 Classification of HPLC Separations by Column Dimensions

Please do leave your comments and suggestions on the topic covered.

Filed Under: HPLC

About Dr. Deepak Bhanot

Dr Deepak Bhanot is a seasoned professional having nearly 30 years expertise
beginning from sales and product support of analytical instruments. After
completing his graduation and post graduation from Delhi University and IIT
Delhi he went on to Loughborough University of Technology, UK for doctorate research in
analytical chemistry. His mission is to develop training programs on analytical techniques and
share his experiences with broad spectrum of users ranging from professionals engaged in
analytical development and research as well as young enthusiasts fresh from academics who
wish to embark upon a career in analytical industry.

Comments

Bamgbola Adeyemi says:

April 3, 2014 at 2:00 am

what is the most appropriate C18 columns for HPLC Analysis of spices using UV
detector and isocratic separation.

Reply

Dr. Deepak Bhanot says:

April 3, 2014 at 9:43 am

Dear Bamgbola,
I would like to o er my suggestions but rst I would like to know what are your
target compounds which you wish to determine in pulses using HPLC. Please feel
free to contact me also on my mail id deepak.bhanot@aurigaresearch.com.
regards

Reply

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