• Mahin Basha
Part of the Springer Protocols Handbooks book series (SPH)


The successful chromatographic separation of analytes in a mixture depends upon the selection of the most appropriate process of chromatography followed by the optimization of the experimental conditions associated with the separation. Optimization requires an understanding of the processes that are occurring during the development and elution and of the calculation of a number of experimental parameters characterizing the behavior of each analyte in the mixture.

In any chromatographic separation, two processes occur concurrently to affect the behavior of each analyte and hence the success of the separation of the analytes from each other. The first involves the basic mechanisms defining the chromatographic process such as adsorption, partition, ion exchange, ion pairing, and molecular exclusion. These mechanisms involve the unique kinetic and thermodynamic processes that characterize the interaction of each analyte with the stationary phase. The second general process defines the other processes, such as diffusion, which tend to oppose the separation and which result in non-ideal behavior of each analyte. This chapter explains about paper and thin-layer chromatography, HPLC, GC, and gel filtration chromatography.

Key words

Paper chromatography Thin-layer chromatography HPLC GC Gel filtration chromatography 



High-performance liquid chromatography


Relative flow


Thin-layer chromatography


  1. 1.
    Rajan K (2011) Analytical techniques in biochemistry and molecular biology. Springer, New York. eBook ISBN978-1-4419-9785-2Google Scholar
  2. 2.
    Wilson K, Walker J (2010) Principles and techniques of biochemistry and molecular biology. Cambridge University Press, Cambridge. ISBN 978-0-521-73167-6CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Mahin Basha
    • 1
  1. 1.P. M. Sayeed Calicut University CentreUniversity of CalicutLakshadweepIndia

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