Simutaneously Determination of Five Active Ingredients in Black Curd Beans Tables by HPLC

OBJECTIVE To establish an HPLC method to simutaneously determine the content of geniposide, soybean glycoside, genistin, soybean aglycone and genistein in Black Curd Beans tables. METHODS Hypersil ODS 2 chromatographic column (200 mm×4.6 mm, 5 μm) was used, the mobile phase was consisted of methyl cyanides, methanol and 1.8% glacial acetic acid as A, B and C, respectively. The elution procedure was 0 min(A: 8%, B: 10%, C: 82%)→28 min(A: 8%, B: 10%, C: 82%)→28.1 min(A: 0%, B: 38%, C: 62%)→55 min(A: 0%, B: 38%, C: 62%), the flow rate was 1 mL·min^-1, the detected wavelength was 254 nm and the temperature of column was 30 ℃. RESULTS The linear relationship of geniposide, soybean glycoside, genistin, soybean aglycone and genistein were 12.36-396.00, 3.50-112.00, 3.36-116.00, 3.00-96.00 and 1.63- 52.00 μg·mL^-1, respectively; the average recoveries were 100.0%, 100.3%, 99.8%, 99.4% and 99.9%, the RSD were 0.14%, 0.97%, 0.83%, 0.98% and 1.23%. CONCLUSION This method is sensitive, specific and reproducible, and can be used to control the quality of this preparation.