超高效液相色谱-串联质谱法测定大豆中异黄酮素含量  

Determination of isoflavone in soybean by ultra performance liquid chromatography-tandem mass spectrometry

作　　者：林胜军[1] 刘国平[1] 薛荣旋 卢丽明[1] 温伊蕾 黄莹偲 LIN Shengjun;LIU Guoping;XUE Rongxuan;LU Liming;WEN Yilei;HUANG Yingsi(Zhongshan Municipal Center for Disease Control and Prevention,Guangdong Zhongshan 528403,China)

机构地区：[1]中山市疾病预防控制中心,广东中山528403

出　　处：《中国食品卫生杂志》2023年第3期374-380,共7页Chinese Journal of Food Hygiene

基　　金：中山市医学科研项目(2018J173)。

摘　　要：目的以染料木黄酮、大豆苷元和黄豆黄素的含量为指标,建立超高效液相色谱-串联质谱测定大豆中异黄酮素总量的方法。方法样品用乙醇-水(3+1,V/V)提取,经过β-葡萄糖苷酶水解后,用Acquity UPLC?BEH C18柱(2.1 mm×100 mm,1.7μm)分离,以甲醇和0.1%甲酸溶液(含5 mmol/L乙酸铵)作为流动相进行梯度洗脱,电喷雾负离子模式(ESI-)电离,多反应监测模式进行检测。结果染料木黄酮、大豆苷元在5.0~500μg/L范围内线性关系良好(r>0.995),方法的检出限分别为2.7、4.0 mg/kg,在300.0、600.0、1200 mg/kg添加水平的平均回收率为89.4%~102.2%,相对标准偏差(n=6)为1.5%~4.6%;黄豆黄素在0.5~50.0μg/L范围内线性关系良好(r>0.995),方法的检出限为0.6 mg/kg,在30.0、60.0、120.0 mg/kg添加水平的平均回收率为85.7%~104.0%,相对标准偏差(n=6)为1.3%~2.8%。结论该方法简单、灵敏、准确可靠,适用于大豆中异黄酮素含量的测定。Objective To establish a method for the determination of isoflavone in soybean by ultra performance liquid chromatography tandem mass spectrometry with the contents of genistein,daidzein and glycitein as the indexes.Methods The samples were extracted by ethanol-water(3+1,V/V),separated by Acquity UPLC®BEH C18 column(3.0 mm×150 mm,1.7μm)and then eluted by methanol and 0.1%formic acid solution(containing 5 mmol/L ammonium acetate)as mobile phase,detected by UPLC-MS/MS under the electrospray ionization(ESI-)anion ion mode with multiple reaction monitoring mode.Results The linear relationship between genistein and daidzein was good in the range of 5.0-500μg/L(r>0.995),the detection limits were 2.7 and 4.0 mg/kg,respectively.The average recoveries at the addition levels of 300.0,600.0,and 1200 mg/kg were 89.4%to 102.2%and the relative standard deviations(n=6)were 1.5%to 4.6%.The linear relationship of glycitein was good in the range of 0.5-50.0μg/L(r>0.995),the detection limit was 0.6 mg/kg.The average recoveries at the addition levels of 30.0,60.0,and 120.0 mg/kg were 85.7%to 104.0%and the relative standard deviations(n=6)were 1.3%to 2.8%.Conclusion The method is simple,sensitive and accurate,which is suitable for the determination of isoflavone in soybean.

关 键 词：染料木黄酮 大豆苷元 黄豆黄素 大豆 异黄酮素 

分 类 号：R155[医药卫生—营养与食品卫生学]