超高效液相色谱-串联质谱法测定鸡肉中5种β-受体激动剂残留量

Determination of 5 kinds of β-agonist residues in chicken 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)同时测定鸡肉中克伦特罗、溴布特罗、溴代克伦特罗、特布他林、沙丁胺醇5种β-受体激动剂残留量的分析方法。方法在鸡肉样品加入β-葡萄糖醛酸酶进行水解,经混合阳离子(MCX)固相萃取柱净化,利用超高效液相色谱-串联质谱法检测,内标法定量。同时考察了不同的酶制剂对鸡肉中β-受体激动剂含量测定的影响。结果 5种β-受体激动剂在0.1～50μg/kg浓度范围内呈现良好的线性关系,相关系数均大于0.9991,方法的检出限(limits of detection,LOD)为0.02～0.24μg/kg,定量限(limits of quantitation,LOQ)为0.1～0.71μg/kg。在0.5、5、40μg/kg添加水平下,5种β-受体激动剂的平均回收率为77.8%～117.5%,相对标准偏差小于7.9%。在相同的条件下IMCSzyme~?比蜗牛β-葡萄糖醛酸酶能够更有效地解离鸡肉中轭合的特布他林和沙丁胺醇,测得的含量提高,结果稳定。结论本方法缩短了样品水解时间,提高了β-受体激动剂的检测效率,适用于同时检测鸡肉中5种β-受体激动剂含量。

Objective To establish a method for simultaneous determination of 5 kinds of β-agonists（clenbuterol, brombuterol, bromchlorbuterol, terbutaline and salbutamol） in chicken muscle by ultra performance liquid chromatography-tandem mass spectrometry（UPLC–MS/MS）. Methods Chicken meat samples were hydrolyzed by β-glucuronidase. And then the extracts were purified and concentrated by MCX column, analyzed with UPLC-MS/MS and quantified by internal standard method. The hydrolysis efficiency of β-agonists in chicken muscle was also evaluated, when chicken meat samples hydrolyzed with different enzymes from different sources. Results The calibration curves were good linear between 0.1-50 μg/kg, the correlation coefficient for all analytes were higher than 0.9991. The limits of detection（LOD） of 5 kinds of β-agonists in chicken meat were 0.02-0.24 μg/kg, and the limits of quantitation（LOQ） were 0.1-0.71 μg/kg. The recoveries were ranged from 77.8% to 117.5% for the 5 kinds of β-agonist residues at 3 spiked levels of 0.5, 5 and 40 μg/kg with relative standard deviations（RSDs） less than 7.9%. Under the same conditions, recombinant β-glucuronidase（IMCSzyme） was able to dissociate terbutaline and salbutamol conjugated in chicken more efficiently than β-glucuronidase from Helix pomatia. The measured content was increased and the results were stable. Conclusion This method shortens the hydrolysis time of the sample, improves the detection efficiency of the β-agonist, which is suitable for simultaneously detecting the content of 5 β-agonists in the chicken.