在线固相萃取-高效液相色谱-串联质谱法测定猪肉和羊肉中10种β-受体激动剂
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摘要
建立在线固相萃取-高效液相色谱-串联质谱检测猪肉及羊肉中10种β-受体激动剂的全自动分析方法。样品于37℃酶解16 h后,经MCX在线固相萃取小柱净化,采用XBridge C_(18)色谱柱分离,以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,在电喷雾电离正离子模式下,采用多反应监测模式检测,内标法定量。结果表明:10种β-受体激动剂在0.01～10.00 ng/mL范围内线性关系良好,相关系数均大于0.999 0;检出限为0.004～0.040μg/kg,定量限为0.02～0.20μg/kg;方法回收率为76.5%～107.7%,相对标准偏差小于10%。该方法分析速度快、灵敏度高,可用于猪肉和羊肉中10种β-受体激动剂的定性及定量分析。
In this research, we developed a fully automated method to determine 10 β-agonists in pork and lamb using on-line solid phase extraction coupled to high performance liquid chromatography-tandem mass spectrometry(HPLC-MS-MS). After being enzymatically digested at 37 ℃ for 16 h, samples were purified by on-line solid phase extraction on an MCX column, and then separated on an XBridge C_(18) column by gradient elution using 0.1% formic acid solution and acetonitrile as the mobile phase. Mass spectrometric data were acquired under multiple reaction monitoring(MRM) mode using positive ion electrospray ionization. Quantitation was performed by the internal standard method. The results showed a good linear relationship in the concentration range of 0.01–10.00 ng/m L was achieved for all analytes with acorrelation coefficient higher than 0.999 0. The limits of detection(LOD) were in the range of 0.004–0.040 μg/kg and the limits of quantitation(LOQ) were in the range of 0.02–0.20 μg/kg. The recoveries of the method ranged from 76.5% to 107.7%, with relative standard deviation(RSD)lower than 10%. These results demonstrate that the method is simple, sensitive, and can be used for quantification of 10 β-agonists in pork and lamb samples.
In this research, we developed a fully automated method to determine 10 β-agonists in pork and lamb using on-line solid phase extraction coupled to high performance liquid chromatography-tandem mass spectrometry(HPLC-MS-MS). After being enzymatically digested at 37 ℃ for 16 h, samples were purified by on-line solid phase extraction on an MCX column, and then separated on an XBridge C_(18) column by gradient elution using 0.1% formic acid solution and acetonitrile as the mobile phase. Mass spectrometric data were acquired under multiple reaction monitoring(MRM) mode using positive ion electrospray ionization. Quantitation was performed by the internal standard method. The results showed a good linear relationship in the concentration range of 0.01–10.00 ng/m L was achieved for all analytes with acorrelation coefficient higher than 0.999 0. The limits of detection(LOD) were in the range of 0.004–0.040 μg/kg and the limits of quantitation(LOQ) were in the range of 0.02–0.20 μg/kg. The recoveries of the method ranged from 76.5% to 107.7%, with relative standard deviation(RSD)lower than 10%. These results demonstrate that the method is simple, sensitive, and can be used for quantification of 10 β-agonists in pork and lamb samples.
引文
[1]刘佳,谢云峰,任丹丹,等.反相液相色谱-串联质谱法检测猪肝中26种β2-受体激动剂类药物残留[J].分析化学,2014,42(10):1482-1492.DOI:10.11895/j.issn.0253-3820.140457.
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[3]张瑞雨,梁孟军,赵丽,等.固相萃取-高效液相色谱-串联质谱法测定食品中β-受体激动剂[J].中国食品卫生杂志,2017,29(2):167-171.DOI:10.13590/j.cjfh.2017.02.011.
[4]SHAO Bing,JIA Xiaofei,ZHANG Jing,et al.Multi-residual analysis of 16β-agonists in pig liver,kidney and muscle by ultra-performance liquid chromatography tandem mass spectrometry[J].Food Chemistry,2009,114(3):1115-1121.DOI:10.1016/j.foodchem.2008.10.063.
[5]ZHANG Wei,WANG Peilong,SU Xiaoou.Current advancement in analysis of β-agonists[J].Tr AC Trends in Analytical Chemistry,2016,85:1-16.DOI:10.1016/j.trac.2016.08.011.
[6]LI T T,CAO J J,LI Z,et al.Broad screening and identification of β-agonists in feed and animal body fluid and tissues using ultra-high performance liquid chromatography-quadrupole-orbitrap high resolution mass spectrometry combined with spectra library search[J].Food Chemistry,2016,192:188-196.DOI:10.1016/j.foodchem.2015.06.104.
[7]王炼,黎源倩.高效液相色谱法测定动物性食品中4种β2-兴奋剂[J].中国卫生检验杂志,2008,18(7):1227-1230.DOI:10.3969/j.issn.1004-8685.2008.07.001.
[8]金雁,姚家彪,姜莉,等.高效液相色谱法同时测定禽肉中的克伦特罗和沙丁胺醇[J].现代仪器,2005(5):34-36.
[9]刘文卫,钮伟民,蒋立凤,等.酶联免疫与气质联用法测定猪尿中的克伦特罗[J].中国卫生检验杂志,2006,16(5):564-565.DOI:10.3969/j.issn.1004-8685.2006.05.025.
[10]陈小锋,李瑞芳,刘曙照.对克伦特罗具有高特异性的酶联免疫吸附分析法研究[J].分析化学,2013,41(6):940-943.DOI:10.3734/SP.J.1096.2013.20387.
[11]田苗.猪组织中10种β-兴奋剂类兽药残留量的气相色谱-质谱法检测[J].分析测试学报,2010,29(7):712-716.DOI:10.3969/j.issn.1004-4957.2010.07.013.
[12]宋永青,郭文萍,赵蓉,等.气相色谱-质谱法测定动物性食品中3种β-激动剂的残留量[J].肉类研究,2007,21(11):45-49.
[13]朱坚,杨景贤,李波,等.气相色谱-质谱法测定肝、肾和肉中β-受体激动剂残留量[J].分析测试学报,2004,23(9):223-225.DOI:10.3969/j.issn.1004-4957.2004.z1.084.
[14]XIONG Lin,GAO Yaqin,LI Weihong,et al.Simple and sensitive monitoring ofβ2-agonist residues in meat by liquid chromatography-tandem mass spectrometry using a Qu ECh ERS with preconcentration as the sample treatment[J].Meat Science,2015,105:96-107.DOI:10.1016/j.meatsci.2015.03.013.
[15]LIN Y P,LEE Y L,HUNG C F,et al.Determination of multiresidue analysis of β-agonists in muscle and viscera using liquid chromatograph/tandem mass spectrometry with Quick,Easy,Cheap,Effective,Rugged,and Safe methodologies[J].Journal of Food and Drug Analysis,2017,25:275-284.DOI:10.1016/j.jfda.2016.06.010.
[16]SUO Decheng,WANG Ruiguo,WANG Peilong,et al.Pseudo template molecularly imprinted polymer for determination of14 kind ofβ-agonists in animal urine by ultra-high-performance liquid chromatography-tandem mass spectrometry[J].Journal o f C h r o m a t o g r a p h y A,2 0 1 7,1 5 2 6:2 3-3 0.D O I:1 0.1 0 1 6/j.chroma.2017.09.076.
[17]GIANNETTI L,FERRETTI G,GALLO V,et al.Analysis of beta-agonist residues in bovine hair:development of a UPLC-MS/MS method and stability study[J].Journal of Chromatography B,2016,1036/1037:76-83.DOI:10.1016/j.jchromb.2016.09.041.
[18]GRESSLER V,FRANZEN A R L,LIMA G J,et al.Development of a readily applied method to quantify ractopamine residue in meat and bone meal by Qu ECh ERS-LC-MS/MS[J].Journal of Chromatography B,2016,1015/1016:192-200.DOI:10.1016/j.jchromb.2016.01.063.
[19]叶培,岳振峰,肖陈贵,等.液相色谱-串联质谱法检测猪肉中28种β2-受体激动剂[J].食品安全质量检测学报,2013,4(3):682-688.
[20]李阳,苏晓鸥,王瑞国,等.超高效液相色谱-串联质谱法同时测定绵羊唾液中14种β-受体激动剂[J].分析化学,2013,41(6):899-904.DOI:10.3724/SP.J.1096.2013.21050.
[21]张燕,陈国,吕燕,等.液相色谱-串联质谱法快速测定猪肉中克伦特罗残留量[J].肉类研究,2016,30(10):30-34.DOI:10.15922/j.cnki.rlyj.2016.10.006.
[22]中华人民共和国上海出入境检验检疫局,上海市疾病预防控制中心,上海水产大学,等.动物源食品中多种β-受体激动剂残留量的测定液相色谱串联质谱法:GB/T 22286—2008[S].北京:中国标准出版社,2008.
[23]中华人民共和国山东出入境检验检疫局,中华人民共和国上海出入境检验检疫局,中国检验检疫科学研究院.进出口动物源食品中克伦特罗、莱克多巴胺、沙丁胺醇和特步他林残留量的测定液相色谱-质谱/质谱法:SN/T 1924—2011[S].北京:中国标准出版社,2011.
[24]中国兽医药品监察所.动物源性食品中β-受体激动剂残留检测液相色谱-串联质谱法:农业部1025号公告-18—2008[S].北京:中国农业出版社,2008.
[25]祝伟霞,杨冀州,郭慧颖,等.阳离子涡流在线固相萃取/液相色谱-串联质谱快速分析猪尿液中16种β-受体激动剂残留[J].分析测试学报,2015,34(7):802-806.DOI:10.3969/j.issn.1004-4957.2015.07.008.
[26]李丹妮,严凤,吴剑平,等.在线净化液相色谱-串联质谱法测定畜禽粪便中7种β2-受体激动剂[J].分析化学,2014,42(12):1797-1803.DOI:10.11895/j.issn.0253-3820.140719.
[27]WANG G N,WU N P,HE X,et al.Magnetic graphene dispersive solid phase extraction-ultra performance liquid chromatography tandem mass spectrometry for determination ofβ-agonists in urine[J].Journal of Chromatography B,2017,1067:18-24.DOI:10.1016/j.jchromb.2017.09.043.
[28]MAURO D,CIARDULLO S,CIVITAREALE C,et al.Development and validation of a multi-residue method for determination of 18β-agonists in bovine urine by UPLC-MS/MS[J].Microchemical Journal,2014,115:70-77.DOI:10.1016/j.microc.2014.02.012.
[29]金玉娥,郭德华,郑烨,等.液质联用仪测定动物源性食品中11种β2-受体激动剂的研究[J].质谱学报,2007,28(4):193-201.DOI:10.3969/j.issn.1004-2997.2007.04.001.
[30]耿宁,卢剑.高效液相色谱-串联质谱法测定肌肉组织中4种兽药残留[J].肉类研究,2017,31(8):39-43.DOI:10.7506/rlyj1001-8123-201708008.
[2]赵晓燕.瘦肉精的危害及生物化学检测方法[J].综述专论,2017,47(1):51-52.DOI:10.15979/j.cnki.cn62-1064/s.2017.01.007.
[3]张瑞雨,梁孟军,赵丽,等.固相萃取-高效液相色谱-串联质谱法测定食品中β-受体激动剂[J].中国食品卫生杂志,2017,29(2):167-171.DOI:10.13590/j.cjfh.2017.02.011.
[4]SHAO Bing,JIA Xiaofei,ZHANG Jing,et al.Multi-residual analysis of 16β-agonists in pig liver,kidney and muscle by ultra-performance liquid chromatography tandem mass spectrometry[J].Food Chemistry,2009,114(3):1115-1121.DOI:10.1016/j.foodchem.2008.10.063.
[5]ZHANG Wei,WANG Peilong,SU Xiaoou.Current advancement in analysis of β-agonists[J].Tr AC Trends in Analytical Chemistry,2016,85:1-16.DOI:10.1016/j.trac.2016.08.011.
[6]LI T T,CAO J J,LI Z,et al.Broad screening and identification of β-agonists in feed and animal body fluid and tissues using ultra-high performance liquid chromatography-quadrupole-orbitrap high resolution mass spectrometry combined with spectra library search[J].Food Chemistry,2016,192:188-196.DOI:10.1016/j.foodchem.2015.06.104.
[7]王炼,黎源倩.高效液相色谱法测定动物性食品中4种β2-兴奋剂[J].中国卫生检验杂志,2008,18(7):1227-1230.DOI:10.3969/j.issn.1004-8685.2008.07.001.
[8]金雁,姚家彪,姜莉,等.高效液相色谱法同时测定禽肉中的克伦特罗和沙丁胺醇[J].现代仪器,2005(5):34-36.
[9]刘文卫,钮伟民,蒋立凤,等.酶联免疫与气质联用法测定猪尿中的克伦特罗[J].中国卫生检验杂志,2006,16(5):564-565.DOI:10.3969/j.issn.1004-8685.2006.05.025.
[10]陈小锋,李瑞芳,刘曙照.对克伦特罗具有高特异性的酶联免疫吸附分析法研究[J].分析化学,2013,41(6):940-943.DOI:10.3734/SP.J.1096.2013.20387.
[11]田苗.猪组织中10种β-兴奋剂类兽药残留量的气相色谱-质谱法检测[J].分析测试学报,2010,29(7):712-716.DOI:10.3969/j.issn.1004-4957.2010.07.013.
[12]宋永青,郭文萍,赵蓉,等.气相色谱-质谱法测定动物性食品中3种β-激动剂的残留量[J].肉类研究,2007,21(11):45-49.
[13]朱坚,杨景贤,李波,等.气相色谱-质谱法测定肝、肾和肉中β-受体激动剂残留量[J].分析测试学报,2004,23(9):223-225.DOI:10.3969/j.issn.1004-4957.2004.z1.084.
[14]XIONG Lin,GAO Yaqin,LI Weihong,et al.Simple and sensitive monitoring ofβ2-agonist residues in meat by liquid chromatography-tandem mass spectrometry using a Qu ECh ERS with preconcentration as the sample treatment[J].Meat Science,2015,105:96-107.DOI:10.1016/j.meatsci.2015.03.013.
[15]LIN Y P,LEE Y L,HUNG C F,et al.Determination of multiresidue analysis of β-agonists in muscle and viscera using liquid chromatograph/tandem mass spectrometry with Quick,Easy,Cheap,Effective,Rugged,and Safe methodologies[J].Journal of Food and Drug Analysis,2017,25:275-284.DOI:10.1016/j.jfda.2016.06.010.
[16]SUO Decheng,WANG Ruiguo,WANG Peilong,et al.Pseudo template molecularly imprinted polymer for determination of14 kind ofβ-agonists in animal urine by ultra-high-performance liquid chromatography-tandem mass spectrometry[J].Journal o f C h r o m a t o g r a p h y A,2 0 1 7,1 5 2 6:2 3-3 0.D O I:1 0.1 0 1 6/j.chroma.2017.09.076.
[17]GIANNETTI L,FERRETTI G,GALLO V,et al.Analysis of beta-agonist residues in bovine hair:development of a UPLC-MS/MS method and stability study[J].Journal of Chromatography B,2016,1036/1037:76-83.DOI:10.1016/j.jchromb.2016.09.041.
[18]GRESSLER V,FRANZEN A R L,LIMA G J,et al.Development of a readily applied method to quantify ractopamine residue in meat and bone meal by Qu ECh ERS-LC-MS/MS[J].Journal of Chromatography B,2016,1015/1016:192-200.DOI:10.1016/j.jchromb.2016.01.063.
[19]叶培,岳振峰,肖陈贵,等.液相色谱-串联质谱法检测猪肉中28种β2-受体激动剂[J].食品安全质量检测学报,2013,4(3):682-688.
[20]李阳,苏晓鸥,王瑞国,等.超高效液相色谱-串联质谱法同时测定绵羊唾液中14种β-受体激动剂[J].分析化学,2013,41(6):899-904.DOI:10.3724/SP.J.1096.2013.21050.
[21]张燕,陈国,吕燕,等.液相色谱-串联质谱法快速测定猪肉中克伦特罗残留量[J].肉类研究,2016,30(10):30-34.DOI:10.15922/j.cnki.rlyj.2016.10.006.
[22]中华人民共和国上海出入境检验检疫局,上海市疾病预防控制中心,上海水产大学,等.动物源食品中多种β-受体激动剂残留量的测定液相色谱串联质谱法:GB/T 22286—2008[S].北京:中国标准出版社,2008.
[23]中华人民共和国山东出入境检验检疫局,中华人民共和国上海出入境检验检疫局,中国检验检疫科学研究院.进出口动物源食品中克伦特罗、莱克多巴胺、沙丁胺醇和特步他林残留量的测定液相色谱-质谱/质谱法:SN/T 1924—2011[S].北京:中国标准出版社,2011.
[24]中国兽医药品监察所.动物源性食品中β-受体激动剂残留检测液相色谱-串联质谱法:农业部1025号公告-18—2008[S].北京:中国农业出版社,2008.
[25]祝伟霞,杨冀州,郭慧颖,等.阳离子涡流在线固相萃取/液相色谱-串联质谱快速分析猪尿液中16种β-受体激动剂残留[J].分析测试学报,2015,34(7):802-806.DOI:10.3969/j.issn.1004-4957.2015.07.008.
[26]李丹妮,严凤,吴剑平,等.在线净化液相色谱-串联质谱法测定畜禽粪便中7种β2-受体激动剂[J].分析化学,2014,42(12):1797-1803.DOI:10.11895/j.issn.0253-3820.140719.
[27]WANG G N,WU N P,HE X,et al.Magnetic graphene dispersive solid phase extraction-ultra performance liquid chromatography tandem mass spectrometry for determination ofβ-agonists in urine[J].Journal of Chromatography B,2017,1067:18-24.DOI:10.1016/j.jchromb.2017.09.043.
[28]MAURO D,CIARDULLO S,CIVITAREALE C,et al.Development and validation of a multi-residue method for determination of 18β-agonists in bovine urine by UPLC-MS/MS[J].Microchemical Journal,2014,115:70-77.DOI:10.1016/j.microc.2014.02.012.
[29]金玉娥,郭德华,郑烨,等.液质联用仪测定动物源性食品中11种β2-受体激动剂的研究[J].质谱学报,2007,28(4):193-201.DOI:10.3969/j.issn.1004-2997.2007.04.001.
[30]耿宁,卢剑.高效液相色谱-串联质谱法测定肌肉组织中4种兽药残留[J].肉类研究,2017,31(8):39-43.DOI:10.7506/rlyj1001-8123-201708008.