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

中文关键词:  β-受体激动剂  β-葡萄糖醛酸酶  水解  超高效液相色谱-串联质谱法  鸡肉

英文关键词:β-agonist  β-glucuronidase  hydrolysis  ultra performance liquid chromatography-tandem mass spectrometry  chicken meat

中文摘要:

      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0.1~50 μg/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0.02~0.24 μg/kg, 定量限(limits of quantitation, LOQ)为0.1~0.71 μg/kg。在0.5、5、40 μg/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。

英文摘要:

      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.