超高效液相色谱-串联质谱法测定畜禽饲料中氯霉素类药物含量  

Determination of chloramphenicols in the livestock and poultry feeds by UPLC-MS/MS

作　　者：张伟[1] 彭麟[1] 江善祥[1] 

机构地区：[1]南京农业大学动物医学院,江苏南京210095

出　　处：《南京农业大学学报》2015年第3期453-458,共6页Journal of Nanjing Agricultural University

基　　金：农业部农业行业标准修订项目(2013-086)

摘　　要：[目的]建立一种畜禽饲料中氯霉素、甲砜霉素和氟苯尼考的超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法,为监控畜禽饲料中违法添加该类药物提供方法指导,保障动物源食品安全。[方法]饲料粉碎后,经碱性乙酸乙酯提取,正己烷除脂,Sep-Pak C18固相萃取柱净化,Waters Acquity UPLC BEH C18色谱柱分离,采用乙腈和水为流动相,并用梯度洗脱来提高对3种药物的分离能力。分别以d5-氯霉素、d3-甲砜霉素和d3-氟苯尼考为内标,采用负离子多反应监测模式,6个特征离子通道(m/z:321→152,257;354→185,290;356→185,336)用于氯霉素、甲砜霉素和氟苯尼考的定性和定量检测。[结果]3种氯霉素类药物在1~20μg·kg-1范围内线性良好,相关系数均大于0.99,方法检测限为0.1μg·kg-1,定量限为0.3μg·kg-1。3个加标水平(1、2和5μg·kg-1)的平均回收率为81.41%~112.88%,日内精密度为1.24%~12.92%,日间精密度为0.64%~13.09%。[结论]试验结果完全符合氯霉素类药物检测的相关法规要求,适用于畜禽饲料中氯霉素类药物的定性定量检测。[ Objectives ] The aim of this experiment was to establish a detection method for chloramphenicol, thiamphenicol, and flor- fenicol by ultra performance liquid chromatography-tandem mass spectrometry in the livestock and poultry feeds,which could provide a methodological guidance to monitor these medicinal additives which were illegally added in the feed and further ensure the safety of animal origin food. [ Methods ] The ground samples were extracted with a mixture of ammonia water/ethyl acetate by liquid-liquid extraction, and the fat was removed by hexane. Sep-Pak C is solid phase extraction columns were used to clean up the impurities from the matrix. The separation step of chloramphenicol, thiamphenicol, and florfenicol was performed on Waters Acquity UPLC system with a column of BEH Cls. The mobile phase was acetonitrile and water, and the gradient elution program was used to improve the separating capacity, ds-chloramphenicol, d3-thiamphenicol and d3-florfenicol were used as the mixed internal standards and drug mo- nitoring mode was negative ion multiple reaction monitoring mode. Six characteristic transition reactions（ m/z：321→152,257;354→185,290 ; 356→185,336） were tested for the confirmation and quantification of chloramphenicol, thiamphenicol and florfenicol simul- taneously. [ Results ] The results showed that the calibration curves were good linear at the concentrations of 1-20μg· kg^-1. The rela- tive coefficients （R2） were greater than 0.99. The limit of detection of ehloramphenicol, thiamphenicol and florfenicol was 0.1 Iμg· kg^-1. and the limit of quantification was 0.3 μg·kg^-1. The average recoveries of 3 drugs at the concentrations of 1,2,5 p^g.kg-t ranged from 81.41% to 112.88%. The intra-day RSD of this method was 1.24% - 12.92%, inter-day RSD was 0.64% - 13.09%. [ Conclu- sions ] The test results were fully in line with the related regulatory requirements and the method could be used to detect chloram- phenicol, thiamphenicol,florfenicol in the livestock and poultry f

关 键 词：超高效液相色谱-串联质谱 氯霉素 甲砜霉素 氟苯尼考 畜禽饲料 

分 类 号：S859.79[农业科学—临床兽医学]