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A new simple and efficient RP-HPLC-UV method was developed for the simultaneous analysis of nine isoflavonoids in Huangqi-Gegen herbal pair. Effective chromatographic separation of these components was achieved on a Kromasil C18 column (4.6×250 mm, i.d.; 5 μm) with gradient elution of methanol and 0.2% formic acid aqueous solution at a flow rate of 1.0 mL/min in 60 min. Detection was performed at 250nm. The method was proved to be linear in the ranges of 81.28-406.4, 670.4-3352, 136.96-684.8, 33.376-166.88, 22.848-114.24, 10.176- 50.88, 31.04-155.2, 44.544-222.72 and 26.56-132.8 ng for the nine isoflavonoid: 3'-hydroxypuerarin, puerarin, daidzin, calycosin-7-O-β-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 1.79, 2.02, 1.44, 1.37, 1.26, 1.81, 1.29, 0.97 and 1.77 % respectively. The developed method was successfully applied for the quantitative analysis of constituents in huangqi gegen herbal pair.

RP-HPLC-UV Method for Simultaneous Determination of Nine Isoflavonoids in Huangqi-Gegen Herbal Pair