同时测定乌骨鸡肉中39种兽药及其代谢物UPLC-MS/MS法的建立与应用

Establishment and Application of a UPLC-MS/MS Method for Simultaneous Determination of 39 Veterinary Drugs and Their Metabolites in Black Bone Chicken

摘要:

【目的】全面排查乌骨鸡兽药残留风险。【方法】结合乌骨鸡临床用药种类，利用低温超声提取、固相萃取前处理技术与超高效液相色谱- 串联质谱（ultra performance liquid chromatography-tandem massspectrometry，UPLC-MS/MS），建立了一种同时测定3 类共39 种兽药及其代谢物的快速检测方法。用Mcllvaine-Na2EDTA 和磷酸盐缓冲液对样品进行低温超声提取，再用Oasis HLB 固相萃取柱净化样品，样品经T3 色谱柱分离后，采用基质匹配外标法进行定量。【结果】39 种兽药及其代谢物在基质匹配外标溶液质量浓度为2.0~500.0 μg/L 时呈现良好的线性关系，相关系数均大于0.990，检出限为2.0 μg/kg，定量限为10.0μg/kg，该方法在加标质量分数为10.0~100.0 μg/kg 时的加标回收率为68%~111%，相对标准偏差（n=5）为0.65%~15.12%。该方法灵敏度高、定性及定量结果可靠，可满足对乌骨鸡肉兽药残留的检测要求。使用该方法对60 份市售乌骨鸡肉进行检测，结果显示，市售乌骨鸡肉中检出了氟喹诺酮类、四环素类和磺胺类兽药，其中2 份样品的恩诺沙星检测结果（153.187、174.979 μg/kg）超出最大残留限量；2 份样品的甲氧苄啶检测结果（55.007、111.391 μg/kg）超出最大残留限量；2 份样品的二甲氧苄啶检测结果（153.779、189.801 μg/kg）超出最大残留限量。【结论】市售乌骨鸡肉中存在一定的磺胺类兽药抗菌增效剂残留风险，需要加强抗菌增效剂在乌骨鸡养殖中的用药监管。

Abstract:

[Objective] This study aims to comprehensively investigate the risk of veterinary drug residues inblack bone chicken. [Method] A method for rapid and simultaneous analysis of 39 veterinary drugs and theirmetabolites in three categories was established based on low-temperature ultrasonic extraction, solid-phaseextraction pretreatment, and ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The samples were extracted by Mcllvaine-Na2EDTA and phosphate buffer via ultrasonictreatment at low temperatures, purified by Oasis HLB solid-phase extraction column, separated by T3chromatographic column, and quantified by the matrix matching external standard method. [Result] The 39veterinary drugs and their metabolites showed good linear relationships in the matrix matching external standardsolutions with the mass concentration range of 2.0~500.0 μg/L, with the correlation coefficients greater than0.990. The limits of detection and quantification were 2.0 μg/kg and 10.0 μg/kg, respectively. The spike recoveryrates of the method ranged from 68% to 111% at fortified mass fractions of 10.0~100.0 μg/kg, and the relativestandard deviation (n=5) ranged from 0.65% to 15.12%. The method had high sensitivity and reliable qualitativeand quantitative results, meeting the requirements for the detection and analysis of veterinary drug residues inblack bone chicken. This method was then used to test 60 samples of black bone chicken sold on the market. Theresults showed that fluoroquinolones, tetracyclines, and sulfonamides were detected in commercial black bonechicken. The mass fraction of enrofloxacin, trimethoprim, and diaveridine exceeded the maximum residue limitsin two (153.187, 174.979 μg/kg), two (55.007, 111.391 μg/kg), and two (153.779, 189.801 μg/kg) samples,respectively. [Conclusion] There is a certain risk of the residues of sulfonamides in black bone chicken on themarket, and it is necessary to strengthen the supervision on use of antibiotics in black bone chicken breeding.