液相色谱-四极杆线形离子阱质谱法快速检测牛奶中磺胺类和喹诺酮类药物残留  

Rapid Detection of Sulfonamides and Quinolones Residues in Milk Using Liquid Chromatography-Quadrupole-Linear Ion Trap Mass Spectrometry

作　　者：刘楚君 谢洁 屈子裕 张莉蕴 张谛 戴新华[2] 方向[2] 江游[2] 江海洋[1] 吴聪明[1] LIU Chujun;XIE Jie;QU Ziyu;ZHANG Liyun;ZHANG Di;DAI Xinhua;FANG Xiang;JIANG You;JIANG Haiyang;WU Congming(National Key Laboratory of Veterinary Public Health and Safety,College of Veterinary Medicine,China Agricultural University,Beijing 100193,China;Technology Innovation Center of Mass Spectrometry for State Market Regulation,Center for Advanced Measurement Science,National Institute of Metrology,Beijing 100029,China)

机构地区：[1]中国农业大学动物医学院兽医公共卫生安全全国重点实验室,北京海淀100193 [2]中国计量科学研究院前沿计量科学中心,国家市场监管技术创新中心(质谱),北京昌平100029

出　　处：《中国兽医杂志》2024年第12期20-29,共10页Chinese Journal of Veterinary Medicine

基　　金：国家重点研发计划(2020YFF01014605)。

摘　　要：为给国产质谱提供检测动物性食品中兽药残留的参考依据,本试验首次基于自主研制液相色谱-四极杆线形离子阱质谱法(LC-Q-LIT),建立了牛奶中磺胺类药物和喹诺酮类药物残留的检测方法。样品前处理采用液液萃取方法,牛奶样品经乙腈提取,正己烷脱脂,取适量提取液,经氮气吹至近干,初始流动相复溶后,进行LC-Q-LIT检测。基质匹配标准曲线按照空白添加的方法进行制备。仪器条件按照色谱峰形及其响应强度进行优化。LC-Q-LIT检测牛奶中磺胺类和喹诺酮类药物的方法学验证指标包括线性范围、检测限和定量限、回收率和精密度。将LC-Q-LIT方法与文献中报道的液相色谱-串联质谱(LC-MS/MS)方法进行对比,并采用LC-Q-LIT方法检测5份实际样品。经优化选择依利特C18色谱柱(4.6 mm×150 mm,5μm)分离目标物;以0.1%甲酸水溶液-甲醇(V/V)为11种磺胺类药物的流动相,0.1%甲酸水溶液-0.1%甲酸乙腈(V/V)为9种喹诺酮类药物的流动相进行梯度洗脱;质谱采用正离子电离(ESI+)和选择反应监测模式。结果显示,在牛奶基质中,11种磺胺类药物在5~500μg/L线性范围内呈现良好线性关系(R^(2)≥0.999),检测限、定量限和添加回收率范围分别为1.0~3.0μg/L、2.5~10.0μg/L和77.39%~119.44%,日内精密度为1.21%~6.04%,日间精密度为2.25%~9.45%;9种喹诺酮类药物在10~450μg/L线性范围内呈现良好线性关系(R^(2)≥0.990),检测限、定量限和添加回收率范围分别为5.0~9.0μg/L、10.0~20.0μg/L和81.97%~118.49%,日内精密度为1.64%~10.94%,日间精密度为2.05%~12.69%。LC-Q-LIT方法的回收率与LC-MS/MS方法相近,可达到仪器准确度要求。5份实际样品经LC-Q-LIT检测,未测出磺胺类药物,测得司帕沙星残留量为0.84μg/L。结果表明,LC-Q-LIT方法操作简单、快速、成本低且灵敏度高,定性和定量准确,适用于牛奶中磺胺类和喹诺酮类药物残留的检测分析。To provide a reference for detecting veterinary drug residues in animal-derived foods using domestic mass spectrometry,this study,for the first time,established a method for detecting sulfonamide and quinolone residues in milk based on the independently developed liquid chromatography-quadrupole-linear ion trap mass spectrometry(LC-Q-LIT)system.The sample preparation involved liquid-liquid extraction,with acetonitrile extraction and n-hexane defatting.The extract was evaporated to near dryness under nitrogen and reconstituted in the initial mobile phase for LC-Q-LIT detection.Matrix-matched calibration curves were prepared by spiking blank samples.Instrument conditions were optimized based on chromatographic peak shape and response intensity.The validation parameters of the LC-Q-LIT method included linear range,limits of detection and quantification,recovery,and precision.The LC-Q-LIT method was compared with the liquid chromatography tandem mass spectrometry(LC-MS/MS)method reported in the literature,and 5 actual samples were analyzed using the LC-Q-LIT method.Optimized separation of the target compounds was achieved using an Elite C18 column(4.6 mm×150 mm,5μm).A gradient elution was performed with 0.1%formic acid-water and methanol(V/V)as the mobile phase for 11 sulfonamides,and 0.1%formic acid-water and 0.1%formic acid-acetonitrile(V/V)for 9 quinolones.Electronspray ionization+(ESI+)and selected reaction monitoring were used for mass spectrometry.The results showed that for the 11 sulfonamides,a good linear relationship was observed in the range of 5-500μg/L(R^(2)≥0.999),with limits of detection,limits of quantification,and recoveries ranging from 1.0-3.0μg/L,2.5-10.0μg/L,and 77.39%-119.44%,respectively.Intra-day relative standard deviation ranged from 1.21%to 6.04%,and inter-day relative standard deviation from 2.25%to 9.45%.For the 9 quinolones,a good linear relationship was observed in the range of 10-450μg/L(R^(2)≥0.990),with limits of detection,limits of quantification,and recoveries ranging fro

关 键 词：牛奶 磺胺类 喹诺酮类 液相色谱-四极杆线形离子阱质谱法(LC-Q-LIT) 

分 类 号：S859[农业科学—临床兽医学] O657[农业科学—兽医学]