Separation Techniques in Biochemistry: Electrophoresis and Chromatographic Methods

| Cerebral Link | Publish by Cerebral Publication Private Limited | 2026 | Volume 3 | Page 112-122 |ISBN: 978-81-689463-6-1 | Book Title: - Text-book of Practical Microbiology | Chapter 8: Separation Techniques in Biochemistry: Electrophoresis and Chromatographic Methods

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Abstract

Separation techniques are fundamental to biochemical research and clinical laboratory investigations, enabling the isolation, identification, characterization, and quantitative analysis of complex biological molecules. This chapter focuses on two major approaches: electrophoresis and chromatographic methods. Electrophoresis separates charged biomolecules according to their size, charge, and molecular characteristics under an applied electric field. Important techniques include agarose gel electrophoresis, polyacrylamide gel electrophoresis (PAGE), SDS-PAGE, isoelectric focusing, and capillary electrophoresis, with applications in the analysis of proteins, nucleic acids, enzymes, and clinical samples. Chromatographic methods separate biochemical components based on their differential interactions between stationary and mobile phases. The principles and applications of paper chromatography, thin-layer chromatography, column chromatography, ion-exchange chromatography, size-exclusion chromatography, affinity chromatography, gas chromatography, and high-performance liquid chromatography (HPLC) are discussed. Emphasis is placed on basic principles, instrumentation, experimental procedures, interpretation of results, troubleshooting, and quality control. Together, electrophoretic and chromatographic techniques provide essential analytical tools for modern biochemistry, molecular biology, clinical diagnostics, pharmaceutical research, and biomedical sciences.