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Updated: May 19, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
Automated derivatization with o-phthalaldehyde for estimation of amino acids in plasma using reversed-phase high
J V Gnanou1, S K Srinivas, A V Kurpad
1Core Laboratory, Institute of Population Health and Clinical Research, St. John's National Academy of Health Sciences, Bangalore, India. justin@divnut.net
Abstract:
A fully automated method for quantitative estimation of plasma amino acids using fluorescence detection of ophthaladehyde/2-mercaptoethanol derivatives of the analytes and their separation by gradient elution reversed-phase HPLC has been described. The method is simple and the three-step gradient elution is suitable for routine analysis of a large number of biological samples due to clear resolution, high degree of precision, accuracy, cost-effectiveness and lack of interference from chemical contaminations. Using this method, 19 amino acids were completely resolved and the within-run coefficients of variation ranged from 2.53 to 10.7% with a mean variation of 5.68%.
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