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目的:建立同时测定红三叶中大豆苷元、染料木素、刺芒柄花素和鹰嘴豆芽素A的高效液相色谱方法。方法:采用Kromasil C18色谱柱(250mm×4.6mm,5μm),以乙腈-0.1%磷酸为流动相进行梯度洗脱,流速1.0mL·min-1,检测波长为248nm和260nm。结果:大豆苷元、染料木素、刺芒柄花素和鹰嘴豆芽素A分别在0.01～217.0,0.04～616.0,0.03～428.6,0.04～581.2μg·mL-1范围内线性关系良好(r≥0.9998);平均加样回收率为96.0%～96.6%,RSD0.9%～1.8%。结论:该方法简便、准确,重现性好。
Objective: To establish a high performance liquid chromatography (HPLC) method for the simultaneous determination of daidzein, genistein, thorononetin and chickpea sp. A in red clover. Methods: Kromasil C18 column (250 mm × 4.6 mm, 5 μm) was used with gradient elution with acetonitrile-0.1% phosphoric acid as the mobile phase. The flow rate was 1.0 mL · min-1. The detection wavelength was 248 nm and 260 nm. Results: The linear relationship between daidzein, genistein, thorn formononetin and chickpea sp. A was in the range of 0.01 ~ 217.0,0.04 ~ 616.0,0.03 ~ 428.6,0.04 ~ 581.2μg · mL-1, respectively ≥0.9998). The average recoveries were 96.0% ~ 96.6% and RSD 0.9% ~ 1.8%. Conclusion: The method is simple, accurate and reproducible.