[Simultaneous determination of six flavonoids in Apocynum venetum by HPLC].

ABSTRACT

OBJECTIVE: To develop a high-performance liquid chromatography (HPLC) method for the simultaneous determination of rutin, hyperoside, isoquercetin, astragulin, quercetin, and kaempferol in Apocynum venetum and its extracts. METHOD: The separation was carried out on a Shim pack ODS (4.6 mm x 250 mm, 5 microm) colum eluted with in mobile phases of water containing 0.2% phosphoric acid and acetonitrile containing 0.2% phosphoric acid in acetonitrile gradient mode. The column temperature was 40 degrees C, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm. RESULT: The good seperation of six flavonoids was achieved within 40 min, with the relative standard deviations (RSD) of intra- and inter-day precision < or = 2.0%. Calibration curves of rutin, hyperoside, isoquercetin, astragulin, quercetin, and kaempferol showed good linear relationship (R2 > 0.999 7, n = 6). The average recoveries of the six flavonoids were within 97.30% - 105.8% (RSD 2.6%). Three batches of A. venetum and 2 batches of its extracts were determined. CONCLUSION: The developed method is simple, accurate, and repeatable, and can be readily used as a powerful tool for the quality control of A. venetum and its extracts.