Reverse-Phase High-Performance Liquid-Chromotographic DAD Method for Simultaneous Determination of Five Isoflavones in Millettia nitida var. hirsutissima

摘要

A reverse-phase high-performance liquid-chromatographic photodiode array detection (DAD) method has been developed and validated for the simultaneous determination of five isoflavones in Millettia nitida var. hirsutissima: hirsutissimiside B, sphaerobioside, lanceolarin, formononetin, and afromosin. The method utilized a C_(18) column (4μ m, 250 × 4.6 mm) with an acetonitrile and aqueous H_3PO_4 gradient and ultraviolet detection at 259 nm. The recovery of the method is 94.1-105.9%, and linearity (r > 0.9996) is obtained for all the isoflavones.