RP-HPLC同时测定葛根异黄酮提取部位中葛根素、大豆苷、染料木苷和大豆苷元的含量

RP-HPLC simultaneous determination the content of puerarin,daidzin,genistin,daidzein in the fraction extracted of pueraria isoflavone

目的:建立控制葛根异黄酮提取部位的定性和定量分析方法。方法:用反相高效液相色谱法(RP-HPLC),Kromasil KR100-5C18(4.6mm×250mm,5μm)柱,流动相为乙腈-0.03mol·L-1醋酸铵,梯度洗脱,流速0.7mL·min-1,检测波长250nm,柱温30℃,对葛根异黄酮部位中的主要成分葛根素、大豆苷、染料木苷、大豆苷元进行定量测定。结果:色谱峰分离情况良好,葛根素、大豆苷、染料木苷、大豆苷元可达到定性、定量要求。葛根素在0.13～0.66μg、大豆苷在0.03～0.14μg、染料木苷在0.03～0.16μg、大豆苷元在0.02～0.12μg范围内,线性良好(r>0.9995),平均回收率为98.1%～99.3%。结论:所建方法准确、简便,重复性好,可用于葛根异黄酮部位中主成分的定性、定量考察。

Abstract Objective ：To develop a quantitative determination of puerarin, daidzin, genistin, daidzein in the fraction extracted of pueraria isoflavone. Methods ：The samples were analyzed on a Kromasil KR100 -5 C18 （4. 6 mm × 250 mm,5 μm） column with acetonitrile-0. 03 mol · L^-1 ammonium acetate as a mobile phase using a gradient elution. The flow rate was O. 7 mL · min^-1, and UV detection wavelength was set at 250 nm. Results： The linear ranges of puerarin,daidzin,genistin,daidzein were 0. 13 -0. 66 μg,0. 03 -0. 14 μg,0. 03 -0. 16 μg,0. 02 -0. 12 μg,respectively （ r 〉 0. 9995 ）. The average recoveries were 98. 1% - 99.3 %. Conclusion： The new method is fast, accu- rate, sensitive, selective, and suitable for use in the quality control of pueraria isoflavone fraction.