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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。
OBJECTIVE: To establish a qualitative and quantitative analysis method for controlling the isoflavones extracted from Pueraria lobata. METHODS: A reversed-phase high performance liquid chromatography (RP-HPLC) was used on a Kromasil KR100-5C18 (4.6 mm×250 mm, 5 μm) column. The mobile phase was acetonitrile-0.03 mol·L-1 ammonium acetate with gradient elution at a flow rate of 0.7. The concentration of puerarin, daidzein, genistin and daidzein in the isoflavones from Pueraria lobata were quantitatively determined at mL·min-1, detection wavelength at 250 nm and column temperature at 30°C. Results: The separation of chromatographic peaks was good, and puerarin, daidzin, genistin, daidzein could meet qualitative and quantitative requirements. Puerarin 0.13 to 0.66 μg, daidzin 0.03 to 0.14 μg, genistin 0.03 to 0.16 μg, daidzein in the range of 0.02 to 0.12 μg, the linearity was good (r>0.9995), and the average recovery was 98.1% ~99.3%. Conclusion: The established method is accurate, simple and reproducible. It can be used for the qualitative and quantitative investigation of the main components in the isoflavones from Pueraria lobata.