超高效液相色谱-串联质谱法同时测定保健食品中15种非法添加降糖化学药物

Simultaneous determination of 15 kinds of illegally added hypoglycemic chemicals in health foods by ultra performance liquid chromatography-tandem mass spectrometry

目的建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)同时测定保健食品中15种非法添加降糖化学药物的方法。方法样品采用乙腈超声提取,采用UPLC-MS/MS进行检测,以Atlantis?dC15 (2.1 mm×150 mm, 3μm)为色谱柱, 0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱程序为0～3min,90%A;3～15min,90%～60%A;15～28min,60%～0%A;28～29min, 0%A; 29～30 min, 0～90%A; 30～31 min, 90%A);体积流量0.3 mL/min,柱温40℃,进样体积5μL。选择ESI离子源,多反应监测模式(multiple reaction monitoring, MRM),通过比较MRM通道中样品峰与对照品峰的分子离子峰,二级碎片离子峰等信息确定添加的化学药。结果 15种化学药物的线性范围宽,相关性好,r≥0.995,相对标准偏差(relative standard deviation, RSD)为1.58%～5.12%(n=6);加标浓度为20、100、200 ng/mL时,回收率为81.0%～120.1%,检出限为0.010～0.120μg/g,定量限在0.034～0.39μg/g。应用该方法对48批样品进行了检测,其中有2批次样品中检出盐酸苯乙双胍和格列本脲。结论本方法专属性强,灵敏度高,适合用于保健食品中非法添加15种降糖化学药物的快速检测。

Objective To establish a method for simultaneous determination of 15 kinds of illegally added hypoglycemic chemicals in health foods by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).Methods The sample was ultrasonically extracted with acetonitrile and detected by UPLC-MS/MS,using Atlantis?dC15(2.1 mm×150 mm,3μm)as the column and 0.1%aqueous formic acid(A)-acetonitrile(B)as mobile phase.The gradient elution program was 0-3 min,90%A;3-15 min,90%-60%A;15-28 min,60%-0%A;28-29 min,0%A;29-30 min,0~90%A;30-31 min,90%A.The volume flow rate was 0.3 mL/min,the column temperature was 40℃,and the injection volume was 5μL.Results The linear range of 15 chemical drugs was wide and the correlation was good,r≥0.995,and the relative standard deviation(RSD)was 1.58%-5.12%(n=6).The recoveries at 3 spiked(20,100,200 ng/mL)were 81.0%-120.1%,the limits of detection were 0.010-0.120μg/g,and the limits of quantification were 0.034-0.39μg/g.The 48 batches of samples were tested by this method,and phenformin hydrochloride and glibenclamide were detected in 2 batches.Conclusion This method has strong specificity and high sensitivity,which is suitable for rapid detection of illegally adding 15 kinds of hypoglycemic chemicals in health foods.