Abstract:

An HPLC method was presented for the simultaneous determination of five isoflavones simultaneously in the
aerial (leaves and stems) parts of Pueraria thomsonii Benth ‘Xiangege No.1’. The chromatography was performed on an
AT·LICHROM C18 column (4.6 mm × 250 mm, 5 μm) with a mobile phase of methanol-water by gradient elution at a flow
rate of 1.0 mL/min, and the detection wavelength was set at 250 nm and the column temperature at 30 ℃. The calibration
curves for puerarin, daidzin, genistin, daidzein and genistein were linear in the range of 5.00–50.00 (r = 0.999 4), 1.00–10.00
(r = 0.999 2), 3.64–36.40 (r = 0.999 9), 0.50–5.00 (r = 0.999 8) and 0.10–1.00 μg/mL (r = 0.999 4), respectively. The
LODs were 0.94, 0.24, 0.43, 0.08 and 0.05 ng/L, and the LOQs were 2.85, 0.73, 1.29, 0.25 and 0.15 ng/L. The method had
good precision, stability and reproducibility, and the mean recoveries for puerarin, daidzin, genistin, daidzein and genistein
were 100.11% (RSD = 1.39%), 99.54% (RSD = 1.47%), 98.06% (RSD = 0.32%), 101.52% (RSD = 1.89%) and 101.05%
(RSD = 0.32%), respectively. In conclusion, this study has proposed a rapid, accurate and simple method which allows the
simultaneous determination of five isoflavones in the aerial parts of Pueraria thomsonii Benth.

Key words: Hu’nan Pueraria thomsonii Benth, high performance liquid chromatography, puerarin, daidzin, genistin, daidzein, genistein