Abstract: A method of reversed-phase high performance liquid chromatography (RP-HPLC) using diode array detection (DAD) was developed for the quantitative determination of 3′-geranyl-5,7,4′-trihydroxyisoflavone (compound 1) and 8,9-dihydroxy-1-methoxy-［6′,6′-dimethylpyrano(2′,3′: 2,3)］pterocarpene (compound 2) in Campylotropis hirtella. The separation and quantification were achieved using an Agilent Zorbax SB-C18 column (250 mm×4.6 mm, 5 μm), and mobile phases of acetonitrile and 0.1% formic acid with gradient elution at a flow rate of 1.0 mL/min and 30 ℃. The calibration curves for compounds 1 and 2 were linear in the ranges of 4.4~13.2 μg and 0.428~1.284 μg, respectively. The recoveries were 99.65% and 99.11% with the relative standard deviations of 1.83% and 2.59% (n=5), respectively. This RP-HPLC-DAD method is rather simple, accurate and convenient. It can be used for the quantitative determination of the active flavonoids in Campylotropis hirtella (Franch.) Schindl.

Key words: Campylotropis hirtella (Franch.) Schindl., isoflavone, medicinal plant, pterocarpene, reversed-phase high performance liquid chromatography (RP-HPLC)