As a part of an investigation of the content of herbal drug preparations and herbal medicinal products, we have investigated tinctures (prepared with 40:60 and 60:40 (%, v/v) ethanolwater) of calendula flower, milk-thistle fruit, and passion flower, which are, respectively, widely used for their anti-inflammatory properties, to treat hepatic injuries, and to treat tension and difficully falling asleep. The aim of this workwas to evaluate the flavonoid content, because flavonoids are the active constituents or markers of these herbal drugs, and to establish the best solvent for the extraction of the constituents. The findings reported herein both confirm the presence of several flavonoids previously identified in these herbal drugs and report the presence of others not previously described and identified here for the first time. In general the flavonol content was highest in 60% tinctures.A rapid, reversed-phase (RP) HPLC assaywas developed and validated and enabled good separation of all classes of flavonoids including flavones, flavonols, flavanonols, and flavanolignans. Aglycones and mona, di, and triglycosides (both Oand C-derivatives) of the flavonoids are also easily and satisfactorily separated. This method is thus proposed for the analysis of other herbal drugs or herbal drug preparations in which flavonoids are the active constituents or markers.

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