UPLC-MS/MS法测定减肥降脂通便三类保健食品中添加的37种药物

Determination of 37 drugs illegally added in weight-losing,lipid-lowering and laxative health foods by UPLC-MS/MS

目的:建立超高效液相色谱-串联质谱法(UPLC-MS/MS)同时测定减肥类、降脂类和通便类保健食品中37种非法添加药物。方法:样品以甲醇为溶剂超声提取后,采用Waters BEH Shield RP18(100 mm×2.1 mm,1.7μm)色谱柱分离,以甲醇-15 mmol·L^(-1)乙酸铵(含0.1%乙酸)为流动相梯度洗脱,流速0.2 m L·min^(-1),柱温35℃。采用电喷雾离子源正、负离子模式、多重反应监测(MRM)方式检测37种药物:阿昔莫司、烟酸、麻黄碱、氯噻嗪、氢氯噻嗪、咖啡因、甲基安非他明、番泻苷B、番泻苷A、氨苯蝶啶、氯噻酮、呋塞米、安非他酮、芬氟拉明、维拉帕米、吲达帕胺、布美他尼、苄氟噻嗪、酚酞、氟西汀、苯扎贝特、N-单去甲基西布曲明、大黄酸、西布曲明、N,N-双去甲基西布曲明、吡沙可啶、舍曲林、芦荟大黄素、氟伐他汀、吉非罗齐、洛伐他汀、辛伐他汀、非诺贝特、大黄酚、大黄素、大黄素甲醚、普罗布考。结果:37种化合物在2.5倍定量限到500倍定量限范围内均呈良好的线性关系,相关系数均大于0.995,检出限均在0.01~5.12ng·m L^(-1)范围内,3个浓度水平平均回收率(n=6)均在87.5%～109.5%范围内,RSD均低于8%,74批保健食品检出9批非法添加阳性样品。结论:本法经方法学验证,可用于减肥、降脂和通便类保健食品中37种非法添加药物的快速筛查和定量测定。

Objective：To establish a UPLC-MS/MS method for simultaneous determination of 37 illegally added drugs in weight-losing,lipid-lowering and laxative health foods.Methods：After ultrasonic extracted by methanol,the sample was separated on a Waters BEH Shield RP18（100 mm×2.1 mm,1.7 μm） column with the gradient elution.The mobile phase consisted of 15 mmol·L^-1 ammonium acetate solution（containing 0.1% acetic acid） and methanol with a flow rate of 0.2 m L·min-1.The column temperature was set at 35 ℃.Electrospray ionization（ESI） source was applied and operated in positive and negative mode.Multiple reactions monitoring（MRM） mode was used to quantify the 37 illegally added drugs：acipimox,nicotinic acid,ephedrine,chlorothiazide,hydrochlorothiazide,caffeine,methamphetamine,sennoside B,sennoside A, triamterene,chlortralidone,furosemide,amfebutamone,fenfluramine,verapamil,indapamide,bumetanide,bendroflumethiazide,phenolphthalein,fluoxetine,benzhabeite,N-monodesmethy ibutramine,rhein,sibutramine,N,N-didesmethyls ibutramine,bisacodyl,sertraline,aloeemodin,fluvastatin,gemfibrozil,lovastatin,simvastatin,fenofibrate,chrysophanol,emodin,physcion,and probucol.Results：The 37 drugs showed good linear relationships in the range of 2.5-500 times of the limit of quantification with the correlation coefficients above 0.995.The detection limits ranged between 0.01-5.12 ng·m L^-1.The average recoveries（n=6） were in the range of 87.5%-109.5% at three spiked levels,and the relative standard deviations were less than 8%.Totally,9 of 74 tested healthy food samples contained illegally added drugs.Conclusion：The established method is proved by methodology validation,which is suitable for rapid screening and quantification of 37 illegally added drugs in weight-losing,lipid-lowering and laxative health foods.