固相萃取-超高效液相色谱-串联质谱法快速测定猪尿中莱克多巴胺、沙丁胺醇、克伦特罗残留量

Rapid determination of ractopamine,salbutamol and clenbuterol residues in pigurineby solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry

为了建立测定猪尿中三种β-受体激动剂莱克多巴胺、沙丁胺醇和克伦特罗残留量的固相萃取-超高效液相色谱-串联质谱检测方法,试验采集20份猪尿样品,通过酸法沉淀蛋白,高速离心去除杂质,用MCX(60 mg/3 mL)固相萃取柱净化,以乙腈和0.1%甲酸水溶液为流动相,经液相色谱柱[Waters ACQUITY UPLC BEH C18(100.0 mm×2.1 mm,1.7μm)]分离,采用电喷雾离子源多反应监测模式进行质谱检测,用内标法定量检测莱克多巴胺、沙丁胺醇、盐酸克伦特罗的含量。建立的方法与原农业部1025号公告—11—2008和1063号公告—3—2008中介绍的两种标准方法进行了比较研究。结果表明:三种β-受体激动剂在0.1~50.0μg/L范围内有良好的线性关系,r2≥0.99,检出限为0.1μg/L,定量限为0.2μg/L;在0.1,0.2,1.0μg/L 3个添加水平下,样品平均回收率在80.9%~113.0%之间,相对标准偏差为4.5%~7.6%。与另外两种方法相比,该方法减少了有机溶剂的使用和检测时间。说明该方法简单、快速、可靠,具有良好的灵敏度和准确性,尤其适合大批量猪尿样品中莱克多巴胺、沙丁胺醇、克伦特罗的快速确证和定量分析。

In order to establish a method for the determination of the ractopamine,salbutamol and clenbuterol residues in pig urine solid phaseextraction-ultra performance liquid chromatography-tandem mass spectrometry(SPE-UPLC-MS/MS),the pig urine samples were deproteinizedprecipitated by acid,were centrifuged at high speed to remove impurities,purified by MCX solid phase extraction column,and were separated with acetonitrile-0.1%formic acid solution as mobile phaseby a Waters ACQUITY UPLC BEH C18(100.0 mm×2.1 mm,1.7μm).The contents of ractopamine,salbutamol and clenbuterolwere quantitatively determined by internal standard method using electrospray ion source multi reaction mode.A comparative study was carried out with other methods of No.1025—18—2008 Bulletin of the Ministry of Agriculture and No.1063—3—2008 Bulletin of University of Agriculture.The results showed that there was a good linear relationship among the threeβ-receptor agonistsin the range of 0.1-50μg/L(r2≥0.99),with a detection limit of 0.1μg/L and aquantitative limit of 0.2μg/L,The average recovery ofβ-agonists was 80.9%-113%with a relative standard deviations of 4.5%-7.6%at the addition levels of 0.1,0.2 and 1.0μg/L.The usage of the orgainc solvent and detection that was reduced as companed with other two methods.The results indicated that the method was simple,rapid,and reliable,and had good sensitivity and accuracy,and was especially suitable for the rapid quantitative detection of ractopamine,salbutamol and clenbuterol residues in large quantities of pig urine samples.