液相色谱串联质谱法测定食用肉类及肝脏中9种β-受体激动剂

Determination of residues of nine β-agonists in edible meat and liver by liquid chromatography tandem mass spectrometry

目的建立高效准确的液相色谱串联质谱法（LC-MS/MS）同时测定猪、牛、羊肉及其肝脏中9种β-受体激动剂（菲洛特罗、异丙喘宁、特布他林、沙丁胺醇、莱克多巴胺、克伦特罗、福马特洛、妥布特罗、苯乙醇胺A）残留的确证方法。方法试样（10.0 g）经β-葡糖醛酸苷肽酶和芳基磺酸酯酶混合酶溶液在37℃酶解过夜,游离出结合态的β-激动剂,用高氯酸溶液提取后,分别经过HLB和MCX小柱2次净化处理,甲醇-0.1%甲酸水（10＋90）复溶,采用d6-莱克多巴胺、d9-克仑特罗和d3-沙丁胺醇同位素作为分析的内标,采用电喷雾串联四级杆质谱进行测定。用该方法对深圳市2012—2013年市售的720份样品进行β-受体激动剂的测定。结果 9种目标β-受体激动剂在1～50μg/L范围内该方法线性关系良好,线性相关系数均〉0.995。在所测样品基质中,9种目标β-受体激动剂的方法定量限均为0.5μg/kg,方法平均回收率为66.2%～119.4%,相对标准偏差为0.36%～10.02%。用所建立方法对720份样品进行检测,共检出9份阳性样品,总检出率为1.25%,其中莱克多巴胺检出率为0.56%（4/720）、克伦特罗检出率为0.42%（3/720）、苯乙醇胺A检出率为0.28%（2/720）。结论该方法快速准确,可用于猪、牛、羊肉及其肝脏中9种β-受体激动剂的检测。

Objective To develop an efficient and accurate method for simultaneous determination of multi-residues of 9 β-agonists in meat and liver by using liquid chromatography-tandem mass spectrome- try （ LC-MS/MS）, and to investigate the residues in porcine, bovine, and ovine muscles and livers collect- ed from local markets in Shenzhen. Methods Samples （ 10. 0 g） were hydrolyzed by β-glucuronidase / sulfatase at 37 ℃ overnight to free boundβ-agonists, extracted by perchloric acid, purified by HLB and MCX cartridges, and reconstituted with a methanol -0. 1% formic acid （ 10 ＋90）. The residues of β-ago- nists were determined by electrospray ionization-trandem mass spectrometry, and quantified by internal standard method by d6-ractopamine, d9-clenbuterol and d3-salbutarnol. Results The linear relationships of the nine target β-agonists were optimal in the range of 0-50μg/L and the linear correlation coefficient was 〉 0. 995. The limits of quantification were 0. 5μg/kg for 9 β-agonists in porcine, bovine, and ovine muscles and livers. Average recoveries were 66. 2% - 119.4% with relative standard deviations of 0. 36% -10. 02%. Nine of 720 samples detected by this method were positive, with a detection rate of 1.25%, of which the detection rates were 0. 56% （4/720） for ractopamine, 0. 42% （3/720） for clenbuterol, and 0. 28% （2/720） for phenylethanolamine A. Conclusion This method proved to be quick and accurate for determination of the residues of 9 β-agonists in porcine, bovine and ovine muscle and liver specimens.