A HIGH-THROUGHPUT UPLC-MS/MS FOR THE SIMULTANEOUS ANALYSIS OF SIX PHYTOESTROGENS FROM GENISTA TINCTORIA EXTRACTS

Abstract

A new rapid, sensitive UPLC method coupled with ESI-MS/MS detection was developed and validated for the simultaneous quantification of six phytoestrogens (daidzin, genistin, genistein, daidzein, formononetin, and coumestrol) from Genista tinctoria L. extracts. An adequate chromatographic separation was achieved with gradient elution and a mobile phase consisting of a mixture of 0.1% acetic acid/methanol (70/30, v/v). The total run time was of 5.5 min. The detection of target compounds was performed in multiple reaction monitoring mode. The method was validated and proved to be linear, accurate, and precise over the range of 10–800 ng/mL. The LLOQ values estimated based on the signal to noise ratio were of 5 ng/mL for each analyte, except for daidzein (LLOQ = 10.78 ng/mL). The elaborated assay shows shorter run time, lower solvent consumption, and higher sensitivity than most of the existing LC methods described in the literature. The method was used to quantify the selected phytoestrogens in Genista tinctoria herba or flos extracts. The major compounds found were genistein and its glycosidic form genistin. The method proved to be adequate for the characterization of plant extracts from the point of view of their phytoestrogen content, a group of active compounds with great therapeutic potential.