HPLC coupled on-line to ESI-MS and a DPPH-based assay for the rapid identification of anti-oxidants in Butea superba

N. Nuengchamnong, C.F. de Jong, B. Bruyneel, W.M.A. Niessen, H. Irth, K. Ingkaninan

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

A reversed-phase HPLC coupled on-line to a radical scavenging detection system and MS/MS was developed in order to combine separation, activity determination and structural identification of anti-oxidants in complex mixtures in one run. The sample was separated by HPLC and the eluate split into two flows. The major portion was fed into an electrospray ionisation MS/MS system, while the minor part was mixed with a free radical, 2,2′-diphenyl-1-picrylhydrazyl (DPPH), and the reaction determined spectrophotometrically. The negative peaks, which indicated the presence of anti-oxidant activity, were monitored by measuring the decrease in absorbance at 517 nm. The developed method was successfully applied to the identification of anti-oxidant compounds in a fraction, obtained by solid-phase extraction, of an extract of a Thai medicinal plant, Butea superba Roxb. The anti-oxidant compounds were separated and identified as procyanidin B2, (-)-epicatechin and procyanidin B5. Copyright © 2005 John Wiley & Sons, Ltd.
Original languageEnglish
Pages (from-to)422-8
JournalPhytochemical Analysis
Volume16
Issue number6
DOIs
Publication statusPublished - 2005

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