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目的:建立同时测定葛根药材中6种主要异黄酮类成分含量的HPLC方法。方法:采用RP-HPLC法,KromasilC18(4.6 mm×250 mm,5μm)柱;流动相:甲醇-0.1%枸橼酸溶液,梯度洗脱;流速1.0 mL·min-1,柱温30℃,检测波长250 nm。结果:3’-羟基葛根素、葛根素、大豆苷、染料木苷、芒柄花苷、大豆苷元的线性范围分别为76.2～1 016,65.8～1 316,85.6～1712,49.98～499.8,21.94～219.4,48.5～485 ng,相关系数(R2)均大于0.999 6,6种成分的平均加样回收率分别为99.73%(RSD 0.6%),100.48%(RSD 1.11%),100.37%(RSD 0.79%),100.28%(RSD 1.07%),99.96%(RSD 1.22%),97.36%(RSD 1.59%)。结论:方法简便快速,重复性良好,结果准确可靠,可用于葛根药材中6种主要异黄酮类成分的含量测定。
Objective: To establish an HPLC method for the simultaneous determination of 6 major isoflavones in Radix Puerariae. Methods: RP-HPLC was performed on a Kromasil C18 column (4.6 mm × 250 mm, 5 μm). The mobile phase consisted of methanol-0.1% citric acid solution and eluted with a gradient of 1.0 mL · min- Wavelength 250 nm. Results: The linear range of 3’-hydroxy puerarin, puerarin, daidzin, genistin, monascus glycoside and daidzein were 76.2-1 016,65.8-1 316,85.6-1712,49.98-499.8, 21.94 ~ 219.4 and 48.5 ~ 485 ng respectively, the correlation coefficients (R2) were all greater than 0.999. The average recoveries of six components were 99.73% (RSD 0.6%), 100.48% (RSD 1.11%) and 100.37% 0.79%), 100.28% (RSD 1.07%), 99.96% (RSD 1.22%), 97.36% (RSD 1.59%). Conclusion: The method is simple, rapid, reproducible and accurate. It can be used to determine the contents of six major isoflavones in Radix Puerariae.