不同前处理方法对猪组织中喹诺酮类兽药残留检测效果对比
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摘要
采用超高效液相色谱-三重四极杆质谱和飞行时间质谱技术,比较EMR-L、Oasis PRi ME HLB和液液萃取法对猪组织中16种喹诺酮类兽药残留检测效果的影响。目标兽药以基质匹配工作曲线进行定量,在1.0~100.0 μg/L范围内线性关系良好,在5、10、20 μg/kg加标量条件下使用超高效液相色谱-串联质谱检测,EMR-L和Oasis PRi ME HLB方法处理的猪组织中加标回收率(71.7%~106.9%、70.7%~118.3%)、相对标准偏差(0.2%~16.8%、0.1%~17.8%)、定量限(0.13~0.56、0.27~1.36 μg/kg)和检出限(0.04~0.23、0.08~0.41 μg/kg)均满足方法学要求。针对不同基质成分样品,采用不同的前处理技术减少假阳性结果的产生,以适用于日益增长的动物源性食品兽药残留检测的需求。本研究采用的2种方法操作简便、灵敏度高、重复性好,满足对猪组织中多兽药残留的定性、定量分析。
This study investigated the influence of different sample pretreatments: enhanced matrix removal-lipid(EMR-L) for lipid, Oasis PRi ME hydrophile lipophilic balance(HLB) and liquid-liquid extraction(LLE) on the assay of 16 quinolone residues in porcine tissues by ultra-high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry(UPLC-TQ-MS/MS) or quadrupole-time-of-flight tandem mass spectrometry(Q-TOF/MS). The target compounds were quantified by the matrix-matched calibration curve. Good linearity was observed for all analytes in the range of 1.0–100.0 μg/L. The recoveries using the EMR-L and Oasis PRi ME HLB methods at spiked concentration levels(5, 10 and 20 μg/kg) were 71.7%–106.9% and 70.7%–118.3% with relative standard deviations of 0.2%–16.8% and 0.1%–17.8%, respectively; the limits of quantification of the two methods were 0.13–0.56 and 0.27–1.36 μg/kg and the limits of detection were 0.04–0.23 and 0.08–0.41 μg/kg, respectively. All these figures of merit met the requirements for the determination of quinolones. In order to avoid false positive results, a suitable sample pretreatment method should be chosen for different matrix samples. The two pretreatment methods used in this study, thanks to their ease of operation, high sensitivity and good repeatability, could be suitable for the qualitative and quantitative analysis of multiple veterinary drug residues in porcine tissues.
This study investigated the influence of different sample pretreatments: enhanced matrix removal-lipid(EMR-L) for lipid, Oasis PRi ME hydrophile lipophilic balance(HLB) and liquid-liquid extraction(LLE) on the assay of 16 quinolone residues in porcine tissues by ultra-high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry(UPLC-TQ-MS/MS) or quadrupole-time-of-flight tandem mass spectrometry(Q-TOF/MS). The target compounds were quantified by the matrix-matched calibration curve. Good linearity was observed for all analytes in the range of 1.0–100.0 μg/L. The recoveries using the EMR-L and Oasis PRi ME HLB methods at spiked concentration levels(5, 10 and 20 μg/kg) were 71.7%–106.9% and 70.7%–118.3% with relative standard deviations of 0.2%–16.8% and 0.1%–17.8%, respectively; the limits of quantification of the two methods were 0.13–0.56 and 0.27–1.36 μg/kg and the limits of detection were 0.04–0.23 and 0.08–0.41 μg/kg, respectively. All these figures of merit met the requirements for the determination of quinolones. In order to avoid false positive results, a suitable sample pretreatment method should be chosen for different matrix samples. The two pretreatment methods used in this study, thanks to their ease of operation, high sensitivity and good repeatability, could be suitable for the qualitative and quantitative analysis of multiple veterinary drug residues in porcine tissues.
引文
[1]林黎,张毅,涂小珂,等.液相色谱-串联质谱法同时测定化妆品中的25种喹诺酮类药物[J].色谱,2015,33(3):275-281.DOI:10.3724/SP.J.1123.2014.10029.
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[4]黄子敬,王晓玲,杨钦沾,等.超高效液相色谱-串联质谱法同时测定畜禽肉中磺胺类、喹诺酮类、硝基咪唑类兽药残留[J].分析试验室,2014,33(10):1184-1188.DOI:10.13595/j.cnki.issn1000-0720.2014.0278.
[5]潘明飞,王俊平,方国臻,等.食品中农兽药残留检测新技术研究进展[J].食品科学,2014,35(15):277-282.DOI:10.7506/spkx1002-6630-201415056.
[6]厉文辉,史亚利,高立红,等.加速溶剂萃取-高效液相色谱-串联质谱法同时检测鱼肉中喹诺酮、磺胺与大环内酯类抗生素[J].分析测试学报,2010,29(10):987-992.DOI:10.3969/j.issn.1004-4957.2010.10.001.
[7]郝杰,姜洁,余建龙,等.固相萃取-超高效液相色谱-串联质谱法同时测定动物源性食品中多种兽药残留[J].食品科学,2017,38(12):266-272.DOI:10.7506/spkx1002-6630-201712041.
[8]Council Regulation(EEC).No.470/2009,laying down community procedures for the establishment of residue limits of pharmacologically active substances in foodstuffs of animal origin,repealing Council Regulation(EEC)No.2377/90 and amending Directive 2001/82/EC of the European Parliament and of the Council and Regulation(EC)No.726/2004 of the European Parliament and of the Council[J].Journal of the European Union,2006,L152:11-22.
[9]United States Food and Drug Administration.CFR title 21-food and drugs,Part 556,Tolerances for residues of new animal drugs in food[EB/OL].http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/cfrsearch.cfm?cfrpart=556.
[10]张元,周伟娥,李绍辉,等.动物源性食品中喹诺酮类残留前处理及分析方法的研究进展[J].食品工业科技,2016,37(5):378-383.DOI:10.13386/j.issn.1002-0306.2016.05.069.
[11]ZHANG Y Q,LIU X,LI X,et al.Rapid screening and quantification of multi-class multi-residue veterinary drugs in royal jelly by ultra performance liquid chromatography coupled to quadrupole timeofflight mass spectrometry[J].Food Control,2016,60:667-676.DOI:10.1016/j.foodcont.2015.09.010.
[12]COSTA F P,CALDAS S S,PRIMEL E G.Comparison of QuEChERSsample preparation methods for the analysis of pesticide residues in canned and fresh peach[J].Food Chemistry,2014,165(3):587-593.DOI:10.1016/j.foodchem.2014.05.099.
[13]STOLKER A A M,RUTGERS P,OOSTERINK E,et al.Comprehensive screening and quantification of veterinary drugs in milk using UPLC-TOF-MS[J].Analytical and Bioanalytical Chemistry,2008,391(6):2309-2322.DOI:10.1007/s00216-008-2168-8.
[14]臧国栋,符靖雯,杨钦沾,等.QuEChERS dSPE EMR-Lipid-LCMS-MS测定鸡蛋中3种聚醚类抗球虫药物残留[J].安徽农业科学,2017,45(32):75-76.DOI:10.13989/j.cnki.0517-6611.2017.32.024.
[15]李磊,李海,畅高婧,等.QuEChERS EMR-Lipid-LC/MS/MS测定8种β-受体激动剂[J].食品研究与开发,2016,37(9):178-182.DOI:10.3969/j.issn.1005-6521.2016.09.042.
[16]王飞,宓捷波,李淑静,等.改良的QuEChERS样本前处理/高效液相色谱-串联质谱法检测猪肉中四环素类兽药的残留[J].分析测试学报,2017,36(2):272-275.DOI:10.3969/j.issn.1004-4957.2017.02.021.
[17]HAN L J,MATARRITA J,SAPOZHNIKOVA Y,et al.Evaluation of a recent product to remove lipids and other matrix co-extractives in the analysis of pesticide residues and environmental contaminants in foods[J].Journal of Chromatography A,2016,1449:17-29.DOI:10.1016/j.chroma.2016.04.052.
[18]ANUMOL T,LEHOTAY S J,STEVENS J,et al.Comparison of veterinary drug residue results in animal tissues by ultrahighperformance liquid chromatography coupled to triple quadrupole or quadrupole-time-of-flight tandem mass spectrometry after different sample preparation methods,including use of a commercial lipid removal product[J].Analytical and Bioanalytical Chemistry,2017,409(10):2639-2653.DOI:10.1007/s00216-017-0208-y.
[19]CHEN D M,YU J,TAO Y F,et al.Qualitative screening of veterinary anti-microbial agents in tissues,milk,and eggs of food-producing animals using liquid chromatography coupled with tandem mass spectrometry[J].Journal of Chromatography B,2016,1017/1018:82-88.DOI:10.1016/j.jchromb.2016.02.037.
[20]HAN R W,ZHANG N,YU Z N,et al.Simultaneous determination of 38 veterinary antibiotic residues in raw milk by UPLC-MS/MS[J].Food Chemistry,2015,181:11 9-126.DOI:10.1016/j.foodchem.2015.02.041.
[21]LI X W,GUO P,SHAN Y W,et al.Determination of 82 veterinary drugs in swine waste lagoon sludge by ultra-high performance liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2017,1499:57-64.DOI:10.1016/j.chroma.2017.03.055.
[22]李琴,陶大利,周红,等.超高效液相-串联质谱法同时测定生鲜乳中45种兽药残留量[J].中国乳品工业,2016,44(1):44-47.DOI:10.3969/j.issn.1001-2230.2016.01.010.
[23]吴炜亮,李晓明,朱文亮,等.飞行时间质谱技术及其在食品安全检测中的应用[J].食品与机械,2015,31(3):236-241.DOI:10.13652/j.issn.1003-5788.2015.03.053.
[24]高馥蝶,赵妍,邵兵,等.超高效液相色谱-四极杆-飞行时间质谱法快速筛查牛奶中的农药和兽药残留[J].色谱,2012,30(6):560-567.DOI:10.3724/SP.J.1123.2012.02021.
[25]张科明,梁飞燕,邓鸣,等.QuEChERS结合液相色谱-串联质谱法快速测定猪肉中多类兽药残留[J].色谱,2016,34(9):860-867.DOI:10.3724/SP.J.1123.2016.06010.
[26]李佩佩,张小军,张帅,等.通过式固相萃取结合超高效液相色谱-四级杆飞行时间质谱筛查鱼肉中多类兽药残留[J].浙江海洋学院学报,2017,36(3):228-234.DOI:10.3969/j.issn.1008-830X.2017.03.008.
[27]曹亚飞,康健,常巧英,等.QuEChERS结合液相色谱-串联质谱法快速测定奶酪中多类兽药残留[J].色谱,2015,33(2):132-139.DOI:10.3724/SP.J.1123.2014.10022.
[28]罗辉泰,谢梦婷,黄晓兰,等.分散固相萃取-高效液相色谱-串联质谱法同时测定畜禽肉中63种兽药残留[J].色谱,2015,33(4):354-362.DOI:10.3724/SP.J.1123.2014.12005.
[29]贡松松,顾欣,曹慧,等.超高效液相色谱-四极杆飞行时间质谱法快速筛查生鲜牛乳中的14种磺胺类药物[J].分析测试学报,2014,33(12):1342-1348.DOI:10.3969/j.issn.1004-4957.2014.12.003.
[30]朱万燕,徐文远,张伟,等.超高效液相色谱-四级杆-飞行时间质谱法同时快速检测水产品中37种兽药残留[J].食品安全质量检测学报,2017,8(2):614-619.DOI:10.3969/j.issn.2095-0381.2017.02.041.
[2]孟哲,石志红,吕运开,等.超高效液相色谱-高分辨四级杆飞行时间质谱法快速筛查乳制品中磺胺类与氟喹诺酮类药物[J].分析化学,2014,42(10):1493-1500.DOI:10.11895/j.issn.0253-3820.140299.
[3]韩合敬,魏秀丽,郝智慧,等.超高效液相色谱-串联质谱法测定中兽药散剂中4种喹诺酮药物[J].黑龙江畜牧兽医,2017(1):282-285.DOI:10.13881/j.cnki.hljxmsy.2017.0191.
[4]黄子敬,王晓玲,杨钦沾,等.超高效液相色谱-串联质谱法同时测定畜禽肉中磺胺类、喹诺酮类、硝基咪唑类兽药残留[J].分析试验室,2014,33(10):1184-1188.DOI:10.13595/j.cnki.issn1000-0720.2014.0278.
[5]潘明飞,王俊平,方国臻,等.食品中农兽药残留检测新技术研究进展[J].食品科学,2014,35(15):277-282.DOI:10.7506/spkx1002-6630-201415056.
[6]厉文辉,史亚利,高立红,等.加速溶剂萃取-高效液相色谱-串联质谱法同时检测鱼肉中喹诺酮、磺胺与大环内酯类抗生素[J].分析测试学报,2010,29(10):987-992.DOI:10.3969/j.issn.1004-4957.2010.10.001.
[7]郝杰,姜洁,余建龙,等.固相萃取-超高效液相色谱-串联质谱法同时测定动物源性食品中多种兽药残留[J].食品科学,2017,38(12):266-272.DOI:10.7506/spkx1002-6630-201712041.
[8]Council Regulation(EEC).No.470/2009,laying down community procedures for the establishment of residue limits of pharmacologically active substances in foodstuffs of animal origin,repealing Council Regulation(EEC)No.2377/90 and amending Directive 2001/82/EC of the European Parliament and of the Council and Regulation(EC)No.726/2004 of the European Parliament and of the Council[J].Journal of the European Union,2006,L152:11-22.
[9]United States Food and Drug Administration.CFR title 21-food and drugs,Part 556,Tolerances for residues of new animal drugs in food[EB/OL].http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/cfrsearch.cfm?cfrpart=556.
[10]张元,周伟娥,李绍辉,等.动物源性食品中喹诺酮类残留前处理及分析方法的研究进展[J].食品工业科技,2016,37(5):378-383.DOI:10.13386/j.issn.1002-0306.2016.05.069.
[11]ZHANG Y Q,LIU X,LI X,et al.Rapid screening and quantification of multi-class multi-residue veterinary drugs in royal jelly by ultra performance liquid chromatography coupled to quadrupole timeofflight mass spectrometry[J].Food Control,2016,60:667-676.DOI:10.1016/j.foodcont.2015.09.010.
[12]COSTA F P,CALDAS S S,PRIMEL E G.Comparison of QuEChERSsample preparation methods for the analysis of pesticide residues in canned and fresh peach[J].Food Chemistry,2014,165(3):587-593.DOI:10.1016/j.foodchem.2014.05.099.
[13]STOLKER A A M,RUTGERS P,OOSTERINK E,et al.Comprehensive screening and quantification of veterinary drugs in milk using UPLC-TOF-MS[J].Analytical and Bioanalytical Chemistry,2008,391(6):2309-2322.DOI:10.1007/s00216-008-2168-8.
[14]臧国栋,符靖雯,杨钦沾,等.QuEChERS dSPE EMR-Lipid-LCMS-MS测定鸡蛋中3种聚醚类抗球虫药物残留[J].安徽农业科学,2017,45(32):75-76.DOI:10.13989/j.cnki.0517-6611.2017.32.024.
[15]李磊,李海,畅高婧,等.QuEChERS EMR-Lipid-LC/MS/MS测定8种β-受体激动剂[J].食品研究与开发,2016,37(9):178-182.DOI:10.3969/j.issn.1005-6521.2016.09.042.
[16]王飞,宓捷波,李淑静,等.改良的QuEChERS样本前处理/高效液相色谱-串联质谱法检测猪肉中四环素类兽药的残留[J].分析测试学报,2017,36(2):272-275.DOI:10.3969/j.issn.1004-4957.2017.02.021.
[17]HAN L J,MATARRITA J,SAPOZHNIKOVA Y,et al.Evaluation of a recent product to remove lipids and other matrix co-extractives in the analysis of pesticide residues and environmental contaminants in foods[J].Journal of Chromatography A,2016,1449:17-29.DOI:10.1016/j.chroma.2016.04.052.
[18]ANUMOL T,LEHOTAY S J,STEVENS J,et al.Comparison of veterinary drug residue results in animal tissues by ultrahighperformance liquid chromatography coupled to triple quadrupole or quadrupole-time-of-flight tandem mass spectrometry after different sample preparation methods,including use of a commercial lipid removal product[J].Analytical and Bioanalytical Chemistry,2017,409(10):2639-2653.DOI:10.1007/s00216-017-0208-y.
[19]CHEN D M,YU J,TAO Y F,et al.Qualitative screening of veterinary anti-microbial agents in tissues,milk,and eggs of food-producing animals using liquid chromatography coupled with tandem mass spectrometry[J].Journal of Chromatography B,2016,1017/1018:82-88.DOI:10.1016/j.jchromb.2016.02.037.
[20]HAN R W,ZHANG N,YU Z N,et al.Simultaneous determination of 38 veterinary antibiotic residues in raw milk by UPLC-MS/MS[J].Food Chemistry,2015,181:11 9-126.DOI:10.1016/j.foodchem.2015.02.041.
[21]LI X W,GUO P,SHAN Y W,et al.Determination of 82 veterinary drugs in swine waste lagoon sludge by ultra-high performance liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2017,1499:57-64.DOI:10.1016/j.chroma.2017.03.055.
[22]李琴,陶大利,周红,等.超高效液相-串联质谱法同时测定生鲜乳中45种兽药残留量[J].中国乳品工业,2016,44(1):44-47.DOI:10.3969/j.issn.1001-2230.2016.01.010.
[23]吴炜亮,李晓明,朱文亮,等.飞行时间质谱技术及其在食品安全检测中的应用[J].食品与机械,2015,31(3):236-241.DOI:10.13652/j.issn.1003-5788.2015.03.053.
[24]高馥蝶,赵妍,邵兵,等.超高效液相色谱-四极杆-飞行时间质谱法快速筛查牛奶中的农药和兽药残留[J].色谱,2012,30(6):560-567.DOI:10.3724/SP.J.1123.2012.02021.
[25]张科明,梁飞燕,邓鸣,等.QuEChERS结合液相色谱-串联质谱法快速测定猪肉中多类兽药残留[J].色谱,2016,34(9):860-867.DOI:10.3724/SP.J.1123.2016.06010.
[26]李佩佩,张小军,张帅,等.通过式固相萃取结合超高效液相色谱-四级杆飞行时间质谱筛查鱼肉中多类兽药残留[J].浙江海洋学院学报,2017,36(3):228-234.DOI:10.3969/j.issn.1008-830X.2017.03.008.
[27]曹亚飞,康健,常巧英,等.QuEChERS结合液相色谱-串联质谱法快速测定奶酪中多类兽药残留[J].色谱,2015,33(2):132-139.DOI:10.3724/SP.J.1123.2014.10022.
[28]罗辉泰,谢梦婷,黄晓兰,等.分散固相萃取-高效液相色谱-串联质谱法同时测定畜禽肉中63种兽药残留[J].色谱,2015,33(4):354-362.DOI:10.3724/SP.J.1123.2014.12005.
[29]贡松松,顾欣,曹慧,等.超高效液相色谱-四极杆飞行时间质谱法快速筛查生鲜牛乳中的14种磺胺类药物[J].分析测试学报,2014,33(12):1342-1348.DOI:10.3969/j.issn.1004-4957.2014.12.003.
[30]朱万燕,徐文远,张伟,等.超高效液相色谱-四级杆-飞行时间质谱法同时快速检测水产品中37种兽药残留[J].食品安全质量检测学报,2017,8(2):614-619.DOI:10.3969/j.issn.2095-0381.2017.02.041.