超高效液相色谱-串联质谱法检测牛奶中硝基咪唑类药物及其代谢物残留
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摘要
目的建立三氯乙酸诱导蛋白沉淀净化-超高效液相色谱-串联质谱快速定量检测牛奶中8种硝基咪唑类药物及其代谢物的方法。方法牛奶样品经三氯乙酸蛋白沉淀后高速离心分层,中间层清液过0.22μm聚四氟乙烯滤膜,Hypersil GOLD C18柱(100 mm×2.1 mm, 1.9μm)分离,电喷雾离子化,选择反应监测(SRM)模式检测,基质匹配内标法定量。结果在0.3～10.0 ng/ml浓度范围内,硝基咪唑类药物及其代谢物呈良好的线性关系,相关系数达到0.998 5以上;以3倍信噪比对应的浓度为检出限,方法检出限可达0.1～0.2μg/kg;在0.5、2.0和5.0μg/kg的加标水平,方法回收率为83.6%～111.8%,日内相对标准偏差为2.7%～9.1%,日间相对标准偏差为1.0%～9.3%。结论该检测方法准确、快速、高效、低成本、易操作,能满足牛奶中硝基咪唑类药物及其代谢物残留高通量检测要求。
Objective A method for the quantitative analysis of eight kinds of nitroimidazoles and their metabolites in milk by ultra-performance liquid chromatography-tandem mass spectrometry coupled with simple protein precipitation was established. Methods One gram milk sample was precipitated by trichloroacetic acid, and then stratified by high-speed centrifugation. The middle layer of the supernatant was filtered through a hydrophilic polytetrafluoroethylene membrane. The analytes were separated on a Hypersil GOLD C18 column(100 mm×2.1 mm, 1.9 μm), and detected in selected reaction monitoring(SRM) mode via positive electrospray ionization. The matrix matching and internal standard method was used for quantification. Results The nitroimidazoles and their metabolites showed good linearity in the range of 0.3-10.0 ng/ml, and correlation coefficients were above 0.998 5. The limits of detection of the nitroimidazoles and their metabolites in milk were between 0.1 and 0.2 μg/kg. The recoveries at spiked levels of 0.5, 2.0 and 5.0 μg/kg were within 83.6%-111.8%, the intra-day relative standard deviation(RSD)were within 2.7%-9.1%, and the inter-day RSD were within 1.0%-9.3%. Conclusion The method was accurate, fast, cheap, easy, and could satisfy the requirements of high-throughput monitoring nitroimidazoles in milk.
Objective A method for the quantitative analysis of eight kinds of nitroimidazoles and their metabolites in milk by ultra-performance liquid chromatography-tandem mass spectrometry coupled with simple protein precipitation was established. Methods One gram milk sample was precipitated by trichloroacetic acid, and then stratified by high-speed centrifugation. The middle layer of the supernatant was filtered through a hydrophilic polytetrafluoroethylene membrane. The analytes were separated on a Hypersil GOLD C18 column(100 mm×2.1 mm, 1.9 μm), and detected in selected reaction monitoring(SRM) mode via positive electrospray ionization. The matrix matching and internal standard method was used for quantification. Results The nitroimidazoles and their metabolites showed good linearity in the range of 0.3-10.0 ng/ml, and correlation coefficients were above 0.998 5. The limits of detection of the nitroimidazoles and their metabolites in milk were between 0.1 and 0.2 μg/kg. The recoveries at spiked levels of 0.5, 2.0 and 5.0 μg/kg were within 83.6%-111.8%, the intra-day relative standard deviation(RSD)were within 2.7%-9.1%, and the inter-day RSD were within 1.0%-9.3%. Conclusion The method was accurate, fast, cheap, easy, and could satisfy the requirements of high-throughput monitoring nitroimidazoles in milk.
引文
[1] 陈瑞春, 艾连峰, 郭春海, 等. 液相色谱同位素稀释串联质谱法测定牛奶和奶粉中3种硝基咪唑类禁用兽药及其代谢物的残留量[J]. 中国食品卫生杂志, 2011, 23(6): 543-549.
[2] 汪纪仓, 王大菊, 杨自军, 等. 硝基咪唑类药物残留检测方法研究进展[J]. 中国兽药杂志, 2007, 41(8): 31-33.
[3] 中华人民共和国农业部. 中华人民共和国农业部公告第235号[EB/OL]. (2002-12-24) [2018-12-20]. http://www.moa.gov.cn/zwllm/nybz/200803/t20080304_1028649.htm.
[4] 张毅, 岳振峰, 蓝芳, 等. 分散固相萃取净化与液相色谱/串联质谱法测定牛奶中8类禁用药物残留[J]. 分析化学, 2012, 40(5): 724-729.
[5] 于敏, 张美娟, 张河霞, 等. 高效液相色谱法测定牛奶中硝基咪唑类残留量[J]. 食品安全质量检测学报, 2018, 9(2): 447-451.
[6] 张弛, 潘家荣, 帅瑞琪, 等. 动物性食品中硝基咪唑类兽药多残留酶联免疫检测方法的建立[J]. 核农学报, 2016, 30(2): 323-331.
[7] 何东, 李秀英, 冼燕萍, 等. 气相色谱-质谱法测定祛痘化妆品中4种硝基咪唑类化合物[J]. 分析测试学报, 2015, 34(8): 911-916.
[8] 方力, 邱凤梅, 余新威, 等. 基质分散固相萃取-液相色谱-串联质谱法检测动物源食品中硝基咪唑类药物及其代谢物[J]. 色谱, 2018, 36(5): 431-438.
[9] 刘永明, 曹彦忠, 李金, 等. 液相色谱-串联质谱法快速测定蜂蜜中3种硝基咪唑类药物残留[J]. 色谱, 2010, 28(6): 596-600.
[10] 祝子铜, 雷美康, 彭芳, 等. 快速溶剂萃取-凝胶渗透色谱净化-LC/MS/MS结合测定蜂花粉中硝基咪唑类药物[J]. 药物分析杂志, 2016, 36(3): 522-529.
[11] 黎翠玉, 吴敏, 严丽娟, 等. 高效液相色谱-串联质谱测定动物源性食品中硝基咪唑类药物及其代谢物残留量[J]. 食品安全质量检测学报, 2012, 3(1): 17-22.
[12] 徐纬, 聂四平, 李军, 等. 甲醛-三氯乙酸沉淀-凯氏定氮法测定牛奶中真蛋白质[J]. 理化检验-化学分册, 2015, 51(1): 31-34.
[13] 王海静, 朱风华. 三氯乙酸法与硫酸铜法测定饲料真蛋白比较[J]. 饲料研究, 2012(11): 69-71.
[14] 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. 牛奶和奶粉中甲硝唑、洛硝哒唑、二甲硝唑及其代谢物残留的测定液相色谱-串联质谱法:GB/T 22982—2008 [S]. 北京: 中国标准出版社, 2008.
[2] 汪纪仓, 王大菊, 杨自军, 等. 硝基咪唑类药物残留检测方法研究进展[J]. 中国兽药杂志, 2007, 41(8): 31-33.
[3] 中华人民共和国农业部. 中华人民共和国农业部公告第235号[EB/OL]. (2002-12-24) [2018-12-20]. http://www.moa.gov.cn/zwllm/nybz/200803/t20080304_1028649.htm.
[4] 张毅, 岳振峰, 蓝芳, 等. 分散固相萃取净化与液相色谱/串联质谱法测定牛奶中8类禁用药物残留[J]. 分析化学, 2012, 40(5): 724-729.
[5] 于敏, 张美娟, 张河霞, 等. 高效液相色谱法测定牛奶中硝基咪唑类残留量[J]. 食品安全质量检测学报, 2018, 9(2): 447-451.
[6] 张弛, 潘家荣, 帅瑞琪, 等. 动物性食品中硝基咪唑类兽药多残留酶联免疫检测方法的建立[J]. 核农学报, 2016, 30(2): 323-331.
[7] 何东, 李秀英, 冼燕萍, 等. 气相色谱-质谱法测定祛痘化妆品中4种硝基咪唑类化合物[J]. 分析测试学报, 2015, 34(8): 911-916.
[8] 方力, 邱凤梅, 余新威, 等. 基质分散固相萃取-液相色谱-串联质谱法检测动物源食品中硝基咪唑类药物及其代谢物[J]. 色谱, 2018, 36(5): 431-438.
[9] 刘永明, 曹彦忠, 李金, 等. 液相色谱-串联质谱法快速测定蜂蜜中3种硝基咪唑类药物残留[J]. 色谱, 2010, 28(6): 596-600.
[10] 祝子铜, 雷美康, 彭芳, 等. 快速溶剂萃取-凝胶渗透色谱净化-LC/MS/MS结合测定蜂花粉中硝基咪唑类药物[J]. 药物分析杂志, 2016, 36(3): 522-529.
[11] 黎翠玉, 吴敏, 严丽娟, 等. 高效液相色谱-串联质谱测定动物源性食品中硝基咪唑类药物及其代谢物残留量[J]. 食品安全质量检测学报, 2012, 3(1): 17-22.
[12] 徐纬, 聂四平, 李军, 等. 甲醛-三氯乙酸沉淀-凯氏定氮法测定牛奶中真蛋白质[J]. 理化检验-化学分册, 2015, 51(1): 31-34.
[13] 王海静, 朱风华. 三氯乙酸法与硫酸铜法测定饲料真蛋白比较[J]. 饲料研究, 2012(11): 69-71.
[14] 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. 牛奶和奶粉中甲硝唑、洛硝哒唑、二甲硝唑及其代谢物残留的测定液相色谱-串联质谱法:GB/T 22982—2008 [S]. 北京: 中国标准出版社, 2008.
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