耿宁+卢剑

摘 要：建立同時测定肌肉组织中新霉素、灰黄霉素、多黏菌素B、喹乙醇及其代谢物3-甲基喹啉-2-羧酸（3-methyl-quinoxaline-2-carboxylic acid，MQCA）的高效液相色谱-串联质谱（high performance liquid chromatography-tandem mass spectrometry，HPLC-MS/MS）分析方法。肌肉组织匀浆均质后使用乙腈-5%甲酸水溶液提取，HLB固相色谱柱净化，HPLC分离，电喷雾离子源正离子模式及多反应监测模式检测，外标法定量。结果表明：4 种兽药在质量浓度1.0～200.0 ?g/L范围内均表现出良好的线性关系，R2均大于0.998 0；新霉素、灰黄霉素和多黏菌素B的方法检出限为5.0 μg/kg，喹乙醇及MQCA为0.5 μg/kg；阴性样品中4 种兽药的加标回收率为82.1%～98.4%，日内相对标准偏差（relative standard deviation，RSD）（n=6）为2.2%～6.7%。该方法前处理过程简单、干扰较少，检测结果准确，适用于肌肉组织中4 种兽药的定性与定量检测。

关键词：高效液相色谱-串联质谱法；固相萃取；肌肉组织；兽药残留

Simultaneous Determination of Four Veterinary Drugs in Edible Muscle Tissues by High Performance

Liquid Chromatography-Ttandem Mass Spectrometry

GENG Ning1，2， LU Jian2，*

（1.Quality Association of Jiangsu Province， Nanjing 210029， China；

2.Jiangsu Product Quality Inspection and Testing Institute， Nanjing 210007， China）

Abstract： A novel， simple and rapid method was established for the simultaneous determination of four veterinary drugs （neomycin， griseofulvin， polymyxin B， olaquindox and its metabolite 3-methyl-quinoxaline-2-carboxylic acid （MQCA）） in muscle tissues of food-producing animals by solid phase extraction （SPE） coupled with high performance liquid chromatography tandem triple quadrupole mass spectrometry （HPLC-MS-MS）. After homogenized， muscle tissues were extracted with a mixture of acetonitrile and 5% formic acid solution and the extract was purified by an HLB solid phase extraction column， separated by HPLC， detected by positive electrospray ionization in multiple reaction monitoring mode （MRM）， and quantified by an external standard method. Under the optimal conditions， the calibration for all five analytes showed a linear relationship in the range of 1.0–200.0 ?g/L with correlation coefficients higher than 0.998 0 and the limit of detection was

5.0 ?g/kg for neomycin， griseofulvin， polymyxin B and 0.5 ?g/kg for olaquindox and MQCA， respectively. The mean recoveries of spiked samples （spiked concentration levels： 20.0， 40.0 and 200.0 ?g/L for neomycin， griseofulvin and polymyxin B； 2.0， 4.0 and 20.0 ?g/L for qlaquindox and its metabolite） were ranged from 82.1% to 98.4% with relative standard deviation （RSD， n = 6） between 2.2% and 6.7%. This method is accurate， simple， rapid and applicable to the detection of veterinary drugs in muscle tissues.

Key words： high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）； solid phase extraction； muscle tissue； veterinary drugsendprint

DOI：10.7506/rlyj1001-8123-201708008

中圖分类号：O657.7 文献标志码：A 文章编号：1001-8123（2017）08-0039-05

随着现代养殖技术的迅速发展，兽药在畜禽养殖过程中的使用比例逐渐增加。兽药不仅能够用于畜禽的杀菌治疗，还有促进生长的作用。然而由于养殖人员的技术和知识水平参差不齐，因此经常出现盲目用药、重复给药、药物乱用、实验性用药等现象，这不仅导致畜禽体内大量的兽药残留，还给食品安全带来了巨大隐患[1-3]。

2017年的3·15晚会针对兽药滥用问题做了专题报道，主要涉及到的兽药包括喹乙醇、新霉素、灰黄霉素和黏菌素等杀菌、促生长类药物。其中喹乙醇是一种新型“瘦肉精”，它可以促进畜禽生长，近年来被广泛用于牛、鸡、猪的饲养[4-6]，但毒理学研究表明，喹乙醇对动物具有致癌、致畸的作用。……