大豆及其制品中12种大豆异黄酮的HPLC及HPLC-MS法测定研究

Determination of 12 soybean isoflavones in soybeans and soy-based foods by high performance liquid chromatography and liquid chromatography-mass spectrometry

目的:建立简便、灵敏、准确的测定大豆及其制品中12种大豆异黄酮的高效液相色谱及液-质联用检测方法,并用于大豆及其制品中大豆异黄酮的检测。方法:大豆及其制品经乙腈/水室温下超声辅助提取后,采用HypersilAA-ODS柱(200mm×2.1mm×5μm),以含0.05%三氟醋酸的乙腈/水梯度洗脱,DAD260nm检测,质谱辅助定性,内标法定量。结果:各组分峰面积和内标峰面积的比值与其相应浓度均呈良好的线性关系,r>0.9990。以豆奶粉为基质,各种大豆异黄酮的加标平均回收率均大于90%,RSD在0.35%～14.5%之间。结论:该法操作简便、快速、结果准确、可靠,适用于实际样品的测定。

Objective：To develop an efficient extractive and analytical method for determination of 12 soybean isoflavones in soybeans and soy -based foods by reversed -phase high performance liquid chromatography and liquid chromatography mass spectrometry. Methods：The method used acetonitrile extraction assisted by ultrasonic in room temperature, with apigenin as interhal standard, and identified by mass spectrometry. Samples extracted in acetonitrile - water were diluted to 50% acetonitrile and directly injected for separation on a Hypersil AA - ODS C18 column （200 mm × 2. 1 mm × 5 μm） using an acetonitrile - 0. 05%trifuoroacetic acid （TFA） gradient, DAD detection at 260 nm and mass determination in ESI （ ＋ ） mode. Results：The results showed that the concentration of isoflavones and the ratios of their areas to internal standard area achieved a nice linear relation respectively. Extraction recoveries were more than 90%, intra - day and inter - day RSD were between 3.5% - 14. 5%. Conclusion：This method is found to be sensitive, fast, and accurate, it can be used for determination of isoflavones in soybeans and soy products.