动物源性肉制品中氯霉素测定方法的优化

Optimization on the Determination of Chloramphenicol in Animal-derived Meat Products

复合式净化体系结合超高效液相色谱-串联质谱技术可对动物源性肉制品中的氯霉素进行精确定量分析。基质经超纯水分散溶解,由乙酸乙酯提取,采用正己烷液液萃取,ProElut PLS-A通过式固相萃取先后对提取液进行净化。目标物经AtlantisT3色谱柱,以乙腈和0.1%氨水溶液为流动相进行梯度分离,在负离子(ESI-)多反应监测(MRM)模式下进行定量分析。基于ISO国际标准化组织和AOAC美国分析化学家协会对实验室数据结果的验证要求,所建立的方法进行实验室内部验证,和国内外10家权威机构的实验室间比对。本方法确保氯霉素在0.5~10.0 ng/mL范围内线性关系良好,线性相关系数(R2)≥0.9986。5种基质不同加标水平的回收率在73.34%~91.23%之间,相对标准偏差(RSD)在0.66%~2.91%之间,回收率稳定、重现性良好。同时,验证数据均通过Mandel、Cochran和Grubbs组合统计模型的检验,10家实验室的结果既满足实验室间测试结果的准确度,也满足实验室内测试结果的精密度。所建立的方法普适性强,灵敏度高,准确性和重复性优良,且操作简易,因此被ISO/TC34/SC6国际标准化组织/食品技术委员会/肉禽蛋鱼及其制品分技术委员会修订为最新的国际标准,已于2021年8月27日在全球发布,为世界各国制定相关检测标准提供参考。

A combined purification pretreatment was developed,coupled with the ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)technique on chloramphenicol detection.The matrixes were dissolved and dispersed by ultrapure water,extracted by ethyl acetate,and cleaned using the hexane purification,followed by the ProElut PLS-A SPE system.The residue targets were achieved on Waters Atiantis T3 column with gradient elution of ultra-pure H2O containing 0.1% ammonium hydroxide and acetonitrile as mobile phases,and detected by UPLC-MS/MS with MRM mode via ESI-ionization.An intra-laboratoryvalidation was tested on this method,while an international ring comparison test involved with 10 famous worldwide laboratories was also conducted.The chloramphenicol revealed good linear ranged from 0.5 to 10.0 ng/mL,while the correlation coefficient(R2)was better than 0.9986.The average recoveries on three levels of 5 matrixes(chicken,duck,bovine,ovine,porcine)ranged from 73.34% to 91.23%,with RSD from 0.66% to 2.91%,exhibiting great recovery and repeatability.The compositestatistical modelsby Mandel,Cochran,and Grubbs magnitudes on the data from 10 laboratories verified that all results satisfied the accuracy requirements of inter-laboratory study and the precision requirement of intra-laboratory study.Statistical analysis showed the new method is universal,precise,and simple,thus it had been approved by the ISO/TC34/SC6 committee as the next generation recommended international method for the chloramphenicol determination.And it has been published in international standardization organization system in August 27,2021,to provide the reference for the global standards committee to formulate relevant detection standards.