Automated derivatization with o-phthalaldehyde for estimation of amino acids in plasma using reversed-phase high performance liquid chromatography.
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%.
Core Laboratory, Institute of Population Health and Clinical Research, St. John's National Academy of Health Sciences, Bangalore, India. justin@divnut.net
J V
JV
Gnanou
S K
SK
Srinivas
A V
AV
Kurpad
D000596:Amino Acids / Q000032:analysis / Q000737:chemistry*; D001331:Automation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003362:Cost-Benefit Analysis; D006801:Humans; D010949:Plasma / Q000378:metabolism; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence / Q000379:methods; D009764:o-Phthalaldehyde / Q000737:chemistry*
D000596:Amino Acids; D009764:o-Phthalaldehyde