建立一种超高效液相色谱串联质谱(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)快速测定禽蛋中恩诺沙星、环丙沙星、氧氟沙星、培氟沙星、诺氟沙星、洛美沙星和沙拉沙星共7种喹诺酮类兽药残留的分析方法。禽蛋样品经体积分数为5%的甲酸-乙腈提取喹诺酮类兽药残留,QuEChERS净化后通过快速溶剂蒸发系统浓缩,采用C18色谱柱分离,以体积分数为0.1%的甲酸和乙腈为流动相进行梯度洗脱,电喷雾离子源电离,多反应监测模式采集信号,基质外标法定量。结果表明,7种喹诺酮类兽药在2.0~100.0 ng/mL线性关系良好(R2＞0.995),方法的检出限和定量限分别为0.3和1.0 μg/kg;添加2、5、10 μg/kg三个浓度水平,平均回收率为77.20%~98.47%,相对标准偏差为1.5%~6.2%(n=6)。该方法简单快捷、准确度好、灵敏度高,适用于禽蛋中喹诺酮类兽药残留的快速确证。
An analytical method for rapid determination of enrofloxacin, ciprofloxacin, ofloxacin, pefloxacin, norfloxacin, lomefloxacin and sarafloxacin in poultry eggs by QuEChERS and ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established. The quinolone residues in poultry eggs were extracted by 5% (V/V) formic acid-acetonitrile mixed solution, purified by QuEChERS and concentrated by a rapid solvent evaporation system, and separated on a C18 column with a mobile phase consisted of 0.1% (V/V) formic acid solution and acetonitrile by gradient elution. The quinolone residues were determined in a multiple reaction monitoring (MRM) mode by the electrospray ionization source (ESI+) and quantified with a matrix external standard method. The results showed that all the seven quinolones had good linearity in the concentration range of 2.0-100.0 ng/mL with the correlation coefficients greater than 0.995. The limits of detection (LOD) and limits of quantitation (LOQ) of the method were 0.3 μg/kg and 1.0 μg/kg, respectively, the average recovery rate was 77.20%-98.47%, and the relative standard deviation was 1.5%-6.2%. This method is easy, accurate and sensitive, which is suitable for rapid detection of quinolone residues in poultry eggs.
[1] 李俊锁,邱月明,王超.兽药残留分析[M].上海:上海科学技术出版社,2002:365-390.
[2] HE Xiuting, DENG Maosheng, WANG Qi, et al.Residues and health risk assessment of quinolones and sulfonamides in cultured fish from Pearl River Delta,China[J].Aquaculture,2016,A58(2):38-46.
[3] 张萍.鸡蛋中抗生素类药物残留的研究进度[J].北方药学,2015,12(4):90-92.
[4] HERNÁNDEZ-ARTESEROS J A, BARBOSA J, COMPAÑÓ R, et al. Analysis of quinolone residues in edible animal products[J].Journal of Chromatography A,2002,945(1-2):1-24.
[5] 洪波,曾春芳,高峰,等.高效液相色谱-紫外法测定水产品中四环素类、喹诺酮类抗生素残留[J]. 湖南农业科学,2013 (11):81-84.
[6] 占春瑞,温志海,卜延刚,等.禽肉中多种喹诺酮类兽药残留量的高效液相色谱测定研究[J].食品科学,2005,26(10):172-176.
[7] 刘平,李安运,陈星斌,等.同时分离与测定禽血中的五种喹诺酮兽药的高效液相色谱法研究[J].广东化工,2011,38(5):211-212.
[8] YAO Kai, ZHANG Wei, YANG Linyan, et al.Determination of 11 quinolones in bovine milk using immunoaffinity stir bar sorptive microextraction and liquid chromatography with fluorescence detection[J].J Chromatogr B, 2015,1003(10):67-73.
[9] 杨朝琳,黄方取,洪华嫦,等.液相色谱-质谱联用法检测禽蛋中 8种氟喹诺酮类兽药残留[J].浙江师范大学学报(自然科学版), 2016, 39(1):53-59.
[10] 钱卓真,朱世超,魏博娟,等.高效液相色谱-串联质谱法测定水产品中 19 种喹诺酮类兽药残留量[J].中国渔业质量与标准, 2012,2(3):68-76.
[11] 李迎东,侯新港,齐颖慧,等.QuEChERS结合高效液相色谱串联质谱法同时测定马铃薯中的噻虫嗪、氟啶虫酰胺及其代谢物[J].农药,2019,58(3):199-203.
[12] 许秀敏,龙朝阳,黄伟雄,等.QuEChERS-超高效液相色谱-串联质谱法同时快速检测果蔬中35种甲氧基丙烯酸酯类和三唑类杀菌剂的残留量[J].食品安全质量检测学报,2019,10(4):227-235.
[13] 吴南村,刘春华,乐渊,等.QuEChERS-超高压液相色谱-串联质谱法检测黑胡椒中6种农药残留[J].分析试验室,2018,37(12):73-77.
[14] 刘畅.改良QuEChERS/超高效液相色谱-串联质谱法同时检测蔬菜中14种喹诺酮类抗生素残留[J].分析科学学报,2019,35(1):130-134.
[15] 方从容,高洁,王雨昕,等.QuEChERS-超高效液相色谱-串联质谱法测定鸡蛋中125种兽药残留[J].色谱, 2018, 36(11):57-69.
[16] 刘艳,汪建明,王敏,等. QuEChERS-HPLC-MS/MS测定猪肉猪肝中苯乙醇胺A[J].中国农业科技导报,2018,20(10):131-137.
[17] 中华人民共和国山东出入境检验检疫局.GB/T 20366 —2006动物源产品中喹诺酮类残留量的测定液相色谱-串联质谱法[S].北京:中国标准出版社,2006.
[18] 中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会.GB/T 21312—2007动物源性食品中14 种喹诺酮药物残留检测方法液相色谱-质谱/质谱法[S].北京:中国标准出版社,2007.
[19] 廖艳华,周劭桓,林文斯,等.超高效液相色谱-串联质谱法测定禽蛋中11 种喹诺酮类抗生素残留的样品前处理方法优化[J].中国卫生检验杂志,2012,28(8):910-914.
[20] 中国合格评定国家认可中心. GB/T 27404—2008实验室质量控制规范食品理化检测[S].北京:中国标准出版社,2008.
