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目的用HPLC法测定怀槐异黄酮水解后苷元芒柄花黄素、阿夫罗摩辛、黄豆黄素的含量。方法经正交试验确定的最佳提取与水解工艺制备供试品溶液,采用高效液相法同时测定水解后3种异黄酮苷元含量。色谱柱:DiamonsilTMC18(250 mm×4.6 mm,5μm),流动相:乙腈-质量分数为0.05%的磷酸水溶液(体积比为35∶65),流速:1.0 mL.min-1,检测波长:260 nm,柱温:35℃。结果芒柄花黄素在5.0~25.2 mg.L-1、阿夫罗摩辛在4.2~21.0 mg.L-1、黄豆黄素在4.8~24.0 mg.L-1时峰面积与质量浓度呈良好的线性关系,相关系数分别为1.000 0、0.999 6、0.999 6。芒柄花黄素、阿夫罗摩辛、黄豆黄素的平均回收率分别为98.43%、97.62%、98.38%,RSD分别为1.04%、0.52%、0.23%。结论HPLC测定方法可用于芒柄花黄素、阿夫罗摩辛、黄豆黄素的含量测定。
Objective To determine the content of aglycone, aphrothosin and glycitein in aglycones after hydrolysis of Huaiqi isoflavone by HPLC. Methods The test solution was prepared by the optimal extraction and hydrolysis process determined by orthogonal test. The content of three kinds of isoflavone aglycones after hydrolysis was determined by HPLC. Column: DiamonsilTM C18 (250 mm×4.6 mm, 5 μm), mobile phase: acetonitrile-0.05% phosphoric acid aqueous solution (volume ratio 35:65), flow rate: 1.0 mL.min-1, detection wavelength: 260 nm Column temperature: 35°C. Results The area and concentration of glycopene at 5.0-25.2 mg.L-1, avirosin at 4.2-21.0 mg.L-1, and gamboylin at 4.8-24.0 mg.L-1 were observed. A good linear relationship with correlation coefficients of 1.000 0, 0.999 6, and 0.999 6. The average recoveries of formononetin, afromusine, and glycitein were 98.43%, 97.62%, and 98.38%, respectively, and the RSDs were 1.04%, 0.52%, and 0.23%, respectively. Conclusion The HPLC method can be used to determine the content of formononetin, anovothine and glycitein.