HPLC determination of genistein,formononetin and biochanin A in Cicer arietinum L.

Objective:To establish an HPLC method for the determination of genistein,formononetin and biochanin A in Cicer arietinum L.Method:A Kromasil C18column(250 mm×4.6 mm,5 μm) was adopted with the mobile phase of acetonitrile-0.1% phosphoric acid(40:60),the flow rate was 1.0 mL·min-1.The wavelength of detection was set at 254 nm and the column temperature was at 25 ℃.Results:The calibration curve was linear over the range of 0.42-2.11 μg for genistein(r=0.9990),0.42-2.09 μg for formononetin(r=0.9992) and 0.41-2.06 μg for biochanin A(r=0.9992).The average recovery(n=3)of genistein was 99.6%(RSD=1.5%),that of formononetin was 99.7%(RSD=1.0%)and that of biochanin A was 99.7%(RSD=1.1%),respectively.Conclusion:The method is simple,economical and accurate with good reproducibility.