Classification of HPLC Separations by Column Dimensions

The classification of HPLC methods is governed by several factors such as :

  • Purpose of separation – identification, quantification or collection of purified fractions
  • Available sample size
  • Detection efficiency and choice of detector

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

  • Microbore HPLC
  • Analytical HPLC
  • Semipreparative HPLC
  • Preparative HPLC

Broadly speaking the classification is based on the following parameters :

Classification  id                       Length Packing size                    

Flow rate

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
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

44-69866_250-100MicroboreColumnLC34LG_360x1000

Microbore HPLC Columns

Microbore columns provide the required answers when faced with challenging analytical situations such as limited quantity of sample, need for low eluent consumption, high sensitivity and detection estimations.. Due to low flow rates microbore columns can be used for mass spectrometry detection in LC/MS applications.

Analytical HPLC

Column

Analytical HPLC Column

Majority of academic and industrial applications involve use of analytical HPLC columns. In such situations sample is available in sufficient quantity (pg-ng levels) to meet the analysis requirements. Objective of analytical separations is qualitative identification followed by quantitative estimations. Common HPLC detectors such as UV, RI, Conductivity, Electrochemical,etc provide satisfactory results in analytical separations.

Semi Preparative HPLC

prep_lc2

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

Ultimatezhiebi

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 significantly 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 efficient and time-saving identification, quantification and separations but it seems unlikely that there will be further changes in the column dimension based classification.

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

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

  1. Bamgbola Adeyemi says:

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

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