On-line identification of phenolic compounds of Trifolium species using HPLC-UV-MS and post-column UV-derivatisation.

In an ongoing search for new active compounds in the field of phytoestrogens, a simple HPLC-UV-MS method has been developed in order to identify phenolic compounds. The study was performed on three different species of Trifolium (Leguminosae), namely Trifolium pratense L., T. pallescens Schreb. and T. alpinum L, collected in Switzerland. The comparison between the dichloromethane extracts revealed that the main aglycones are present in the three species whereas the methanolic extracts show different glycosides and malonate derivatives. The compounds of interest were mainly flavonoids, isoflavonoids and clovamides. Their identities were confirmed from retention times, UV and MS analyses and UV shifts following post-column derivatisation.