Specific Proline and Hydroxyproline Detection Method by Post-column Derivatization for High-performance Liquid Chromatography

Abstract

A specific detection method for hydroxyproline by post-column derivatization with 7-chloro-4-nitrobenzo-2-oxa-1, 3-diazole was developed after chiral resolution by ligand-exchange chromatography. The addition of ethylenediaminetetraacetic acid to the reaction mixture was effective to achieve the post-column reaction. The calibration curve for trans-4-hydroxy-L-proline shows linearity from 7. 6pmol up to 7600pmol.