人尿中异黄酮的高效液相色谱分析

Determination of Isoflavones in Human Urine by High Performance Liquid Chromatography

建立了测定人尿中异黄酮组分(大豆苷原、黄豆黄素、雌马酚、染料木黄酮)含量的反相高效液相色谱法。在尿样中加入黄酮作为内标,异黄酮经酶解后在pH=7.0中性条件下用乙酸乙酯(1∶1)提取,然后用0.02%TFA-M eOH-ACN三元梯度洗脱的方法分离异黄酮。在该条件下,大豆苷原、黄豆黄素、雌马酚、染料木黄酮的检出限分别为12.9 nmol/L、13.9 nmol/L、71.6 nmol/L和11.8 nmol/L;回收率均在89%以上。本方法具有测试步骤简单、准确度高、重现性好等优点,适合大批量样品测定。

A reversed phase high performance liquid chromatography method （RP-HPLC） was established for the determination of isoflavones （daidzein, glycitein, equol, genistein） in human urine. After addition of flavone as internal standarad and enzymatic hydrolysis in urine sample, the isoflavones were extracted with acetic ester （ 1 ： 1 ） in medium of pH 7.0, and then were separated by gradient elution with a ternary eluant of 0. 02% trifluoroacetic-CH3OH-acetonitrile. The detection limits were 12.9 nmol/L, 13.9 nmol/L, 71.6 nmol/L and 11.8 nmol/L for daidzein, glycitein, equol and genistein, respectively. The recovery of the isoflavones was over 89%. This proposed method is easier to operate, accurate and reproducible, and suitable for the analysis of great mass of samples.