[Simultaneous determination of nine isoflavonoids in Huangqi Gegen decoction by high performance liquid
Zhiping Wang1, Weimin Li, Ying Gao
1School of Chinese materia medica, Guangzhou University of Chinese Medicine, Guangzhou 510006, China. wzping_jshb@126.com
Objective:
To develop a reversed-phase high performance liquid chromatographic method (RP-HPLC) for the simultaneous analysis of nine isoflavonoids in Huangqi Gegen decoction.
Method:
HPLC determination was performed on a Kromasil C18 column(4.6 mm x 250 mm, 5 microm) and detected at 250 nm. The mobile phase was consisted of methanol and 0.1% citric acid solution with gradient elution. Flow-rate was 1.0 mL x min(-1), and column temperature was 25 degrees C.
Result:
The method was proved to be linear in the ranges of 81.28-406.4, 670.4-3 352, 136.96-684. 8, 33.376-166.88, 22.848-114.24, 10.176-50.88, 31.04-155.2, 44.544-222.72, 26.56-132.8 ng for the nine isoflavonoid, 3'-hydroxypuerarin, puerarin, daidzin, calycosin-7-O-beta-D-glucoside, genistin, ononin, daidzein, calycosin and formononetin respectively. The average recoveries were 99.27%, 102.38%, 98.46%, 103.06%, 101.29%, 99.71%, 102.28%, 97.89% and 100.78%, respectively, RSD was 1.8%, 2.0%, 1.4%, 1.4%, 1.3%, 1.8%, 1.3%, 0.97% and 1.8%, respectively.
Conclusion:
Results obtained show that it's a convenient, accurate and reliable method. The method has been successfully applied to the simultaneous determination of the nine isoflavonoids in Huangqi Gegen decoction.