超高效液相色谱-串联质谱法测定大豆中3种异黄酮素

Determination for 3 kinds of isoflavones in soybean by ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  金雀异黄素  大豆苷元  黄豆黄素  大豆  异黄酮素

英文关键词:genistein  daidzein  glycitein  soybean  isoflavones

中文摘要:

      目的 建立超高效液相色谱-串联质谱法测定大豆中金雀异黄素(genistein)、大豆苷元(daidzein)和黄豆黄素(glycitein)的含量。方法 样品经过乙醇-水(3:1, V:V)提取, 用Acquity UPLC? HSS T3柱(3.0 mm× 150 mm, 1.8 μm)分离, 以甲醇和0.1%甲酸溶液作为流动相进行梯度洗脱, 电喷雾负离子模式(electrospray ionization, ESI-)电离, 多反应监测模式(multiple reaction monitoring mode, MRM)进行检测。结果 金雀异黄素、大豆苷元和黄豆黄素在2.0~200 μg/L范围内线性关系良好(r2>0.999), 方法的检出限分别为0.5、0.4、0.1 mg/kg, 在2.5、10.0、40.0 mg/kg添加水平的平均回收率为79.3%~107%, 相对标准偏差(n=6)为1.3%~6.8%。结论 方法简单、灵敏、准确可靠, 适用于大豆中金雀异黄素、大豆苷元和黄豆黄素等3种异黄酮素含量的测定。

英文摘要:

      Objective To establish a method for determination of genistein, daidzein, glycitein in soybean by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Method The target in the samples was extracted by ethanol-water(3:1, V:V), separated by Acquity UPLC? HSS T3 column (3.0 mm ×150 mm, 1.8 μm) with methanol and 0.1% formic acid solution as mobile phase for gradient elution, then detected by UPLC-MS/MS under the electrospray ionization (ESI-) anion ion mode with multiple reaction monitoring mode (MRM). Result The results showed that genistein, daidzein and glycitein had good linearity in the range of 2.0– 200 μg/L (r2>0.999), the detection limits of the method were 0.5, 0.4, 0.1 mg/kg, respectively, and the average recovery rates at the addition levels of 2.5, 10.0, and 40.0 mg/kg were 79.3%–107%, while the relative standard deviations (n=6) were 1.3% to 6.8%. Conclusion This method is simple, sensitive and accurate, which is suitable for the determination of isoflavones including genistein, daidzein and glycitein from soybean.