Simultaneous determination of two biflavones in Biyanling Tablets by HPLC

Objective of the present study is to develop analytical methods for simultaneous determination of two major biflavones, amentoflavone and heveaflavone, in Biyanling Tablets, thereby providing methodological reference for quality control of anticancer drug Biyanling Tablets. Samples are chromatographed on Diamonsil C18 (4.6 mm × 250 mm, 5 μm) column with a mobile phase of acetonitrile (B)–0.5% acetic acid solution (A); other chromatographic conditions are as follows: elution gradient 0~4 min, B: 35%→45%; 4~20 min, B: 45%→50%, detection wavelength 270 nm, flow rate 1.0 mL/min, and column temperature 30°C. Amentoflavone and heveaflavone show good linear relationships (r≥0.9998 and 0.9996) within a range of 1.56~100 μg/mL, average recoveries are 98.2% and 96.5%, with RSDs of 2.1% and 2.4 % (n = 6), respectively. The method is simple, fast and reproducible, which provides a quantitative analytical method for quality control of Biyanling Tablets.