高效液相色谱-串联质谱法同时测定鸡肉、鸡蛋中喹乙醇和卡巴氧及代谢物残留
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摘要
目的建立高效液相色谱-电喷雾电离串联质谱法测定鸡肉、鸡蛋中喹乙醇、卡巴氧及其代谢物残留量的分析方法。方法组织样品中的卡巴氧和喹乙醇用乙腈-乙酸乙酯(1:1, V:V)溶液提取;代谢物的提取用蛋白酶酶解后经盐酸溶液酸化,采用阴离子交换固相萃取柱净化和富集。分析样品以0.1%甲酸溶液-甲醇进行梯度洗脱,经Waters Xbridge C_(18)色谱柱分离,以液相色谱串联质谱多反应监测(multiple reaction monitoring,MRM)正离子模式进行检测。结果 5种喹喔啉类药物在0.5～50ng/mL浓度范围内线性良好,相关系数r~2为0.9956～0.9988;该方法的检出限和定量限分别为0.1～0.5μg/kg和1.0～2.0μg/kg;鸡蛋和鸡肉样品中,喹乙醇、卡巴氧、脱氧卡巴氧、喹喔啉-2-羧酸和3-甲基喹喔啉-2-羧酸在0.2～2.0μg/kg加标水平的平均回收率为85.1%～108.2%,日内精密度为4.9%～8.7%,日间精密度为6.2%～11.7%。结论该方法简单有效、回收率好、灵敏度高、特异性好,能满足日常检测的需求。
Objective To establish a method for the simultaneous identification and determination of residues of carbadox, olaquindox and related metabolites in chicken and egg by high performance liquid chromatography-tandem mass spectrometry with electrospray ionization. Methods Carboxyl and olaquindox from tissue samples were extracted with acetonitrile-ethyl acetate(1:1, V:V) solution. The metabolites were acidized by hydrochloric acid after protease enzymolysis, and then cleaned up with anion solid-phase extraction cartridge. The separation of compounds were carried on a Waters Xbridge C_(18) column by gradient elution using acetonitrile-0.1% formic acid as mobile phase, and analyzed by high performance liquid chromatography-tandem mass spectrometry in the positive electrospray ionization under multiple reaction monitoring mode(MRM). Results The 5 quinolines had good linear relationships in the range of 0.5-50 ng/mL, and the correlation coefficients were 0.9956-0.9988. The limits of detection and the limits of quantification were 0.1-0.5 μg/kg and 1.0-2.0 μg/kg, respectively. The average recoveries of olaquindox, carbadox, desoxycarbadox, quinoxalie-2-carboxylic acid, and methyl-3-quinoxaline-2-carboxylic acid in egg and chicken samples of 0.2-2.0 μg/kg were 85.1%-108.2%, and intra-day precision and inter-day precision were 4.9%-8.7% and 6.2%-11.7%, respectively. Conclusion This method is simple and effective, with good recovery, high sensitivity and specificity, which can meet the needs of daily detection.
Objective To establish a method for the simultaneous identification and determination of residues of carbadox, olaquindox and related metabolites in chicken and egg by high performance liquid chromatography-tandem mass spectrometry with electrospray ionization. Methods Carboxyl and olaquindox from tissue samples were extracted with acetonitrile-ethyl acetate(1:1, V:V) solution. The metabolites were acidized by hydrochloric acid after protease enzymolysis, and then cleaned up with anion solid-phase extraction cartridge. The separation of compounds were carried on a Waters Xbridge C_(18) column by gradient elution using acetonitrile-0.1% formic acid as mobile phase, and analyzed by high performance liquid chromatography-tandem mass spectrometry in the positive electrospray ionization under multiple reaction monitoring mode(MRM). Results The 5 quinolines had good linear relationships in the range of 0.5-50 ng/mL, and the correlation coefficients were 0.9956-0.9988. The limits of detection and the limits of quantification were 0.1-0.5 μg/kg and 1.0-2.0 μg/kg, respectively. The average recoveries of olaquindox, carbadox, desoxycarbadox, quinoxalie-2-carboxylic acid, and methyl-3-quinoxaline-2-carboxylic acid in egg and chicken samples of 0.2-2.0 μg/kg were 85.1%-108.2%, and intra-day precision and inter-day precision were 4.9%-8.7% and 6.2%-11.7%, respectively. Conclusion This method is simple and effective, with good recovery, high sensitivity and specificity, which can meet the needs of daily detection.
引文
[1]陈杖榴.兽医药理学[M].北京:中国农业出版社,2001.Chen ZL.Veterinary pharmacology[M].Beijing:China Agriculture press,2001.
[2]World Health organization.Toxicological evaluation of certain veterinary drug residues in food[S].
[3]张秀珍,刘云,徐英江,等.新型喹喔啉类药物在水产养殖中的有效性及安全性研究进展[J].食品安全质量检测学报,2013,4(1):38-44.Zhang XZ,Liu Y,Xu YJ,et al.Effectiveness and security of the new type of quinoxalines in aquaculture[J].J Food Saf Qual,2013,4(1):38-44.
[4]Song J,Qiao X,Chen H.Molecularly imprinted solid-phase extraction combined with high-performance liquid chromatography for analysis of trace olaquindox residues in chick feeds[J].Sci Food Agric,2011,(91):2378-2385.
[5]陆桂萍,曲斌,耿士伟,等.液相色谱-串联高分辨质谱测定鸡肉中卡巴氧、喹乙醇的代谢物残留[J].中国家禽,2016,38(2):33-36.Lu GP,Qu B,Geng SW,et al.Determination of metabolites of carbadox and olaquindox in chicken by LC-HRMS[J].China Poult,2016,38(2):33-36.
[6]Li E,Zhang M,Ma H,et al.Determination of3-methylquinoxaline-2-carboxylic acid of olaquindox marker residue in chicken muscles by liquid chromatography-tandem mass spectrometry[J].Chin J Chromatogr,2018,36(5):446-451.
[7]李二粉,张媚玉,马合勤,等.液相色谱-串联质谱测定鸡肉中喹乙醇残留标示物3-甲基喹恶啉-2-羧酸[J].色谱,2018,36(5):446-451.Li EF,Zhang MY,Ma HQ,et al.Determination of3-methylquinoxaline-2-carboxylic acid of olaquindox marker residue in chicken muscles by liquid chromatography-tandem mass spectrometry[J].Chin J Chromatogr,2018,36(5):446-451.
[8]欧阳姗,庞国芳,谢丽琪,等.动物组织中卡巴氧和喹乙醇以及相关代谢产物的液相色谱-串联质谱检测方法[J].分析测试学报,2008,27(6):590-594.Ou YN,Pang GF,Xie LQ,et al.Determination of the residues of carbadox,olaquindox and related metabolites in bovine and porcine muscle and liver tissues by LC-MS/MS[J].J Instrum Anal,2008,27(6):590-594.
[9]贝亦江,王扬,何丰,等.高效液相色谱法测定水产品中喹乙醇代谢物残留[J].食品科学,2013,34(10):255-258.Bei YJ,Wang Y,He F,et al.Determination of olaquindox metabolite residues in fishery products by high performance liquid chromatography[J].Food Sci,2013,34(10):255-258.
[10]赵冬艳,胡冬冬,余伟伟,等.分子印迹固相萃取-高效液相色谱联用检测饲料中喹乙醇的研究[J].粮食与饲料工业,2016,(1):72-75.Zhao DY,Hu DD,Yu WW,et al.Determination of olaquindox in feed using MISPE-HPLC[J].Cerel Feed Ind,2016,(1):72-75.
[11]Sniegocki T.Determination of carbadox and olaquindox metabolites in swine muscle by liquid chromatography/mass spectrometry[J].JChromatogr B,2014,(944):25-29.
[12]Wu Y,Yu H,Wang Y,et al.Development of a high-performance liquid chromatography method for the simultaneous quantification of quinoxaline-2-carboxylic acid and methyl-3-quinoxaline-2-carboxylic acid in animal tissues[J].J Chromatogr A,2007,1146(1):1-7.
[13]郑玲,吴玉杰,李湧,等.高效液相色谱-串联质谱法测定动物源食品中3-甲基喹喔啉-2-羧酸和喹喔啉-2-羧酸残留[J].色谱,2012,30(7):660-664.Zhen L,Wu YJ,Li Y,et al.Determination of the residues of3-methyl-quinoxaline2-carboxylic acid and quinoxaline-2-carboxylic acid in animal origin foods by high performance liquid chromatographytandem mass spectrometry[J].Chin J Chromatogr,2012,30(7):660-664.
[14]郑玲,吴玉杰,赵永锋,等.高效液相色谱-串联质谱法测定动物源食品中喹乙醇与卡巴氧残留量[J].分析测试学报,2014,33(1):21-26.Zhen L,Wu YJ,Zhao YF,et al.Determination of olaquindox and carbadox residues in animal originated food by high performance liquid chromatography-tandem mass spectrometry[J].J Instrum Anal,2014,33(1):21-26.
[15]Zhang H,Qu W,Tao Y,et al.A convenient and sensitive LC-MS/MSmethod for simultaneous determination of carbadox-and olaquindox-related residues in swine muscle and liver tissues[J].J Anal Methods Chem,2018,(2018):1-9.
[16]张静余,杨卫军,严敏鸣.液相色谱-串联质谱法测定水产品中喹乙醇和卡巴氧的代谢物残留量[J].食品安全质量检测学报,2018,9(14):3788-3793.Zhang JY,Yang WJ,Yan MM.Determination of olaquindox and carbadox metabolite residues in aquatic products by liquid chromatography-tandem mass spectrometry[J].J Food Saf Qual,2018,9(14):3788-3793.
[2]World Health organization.Toxicological evaluation of certain veterinary drug residues in food[S].
[3]张秀珍,刘云,徐英江,等.新型喹喔啉类药物在水产养殖中的有效性及安全性研究进展[J].食品安全质量检测学报,2013,4(1):38-44.Zhang XZ,Liu Y,Xu YJ,et al.Effectiveness and security of the new type of quinoxalines in aquaculture[J].J Food Saf Qual,2013,4(1):38-44.
[4]Song J,Qiao X,Chen H.Molecularly imprinted solid-phase extraction combined with high-performance liquid chromatography for analysis of trace olaquindox residues in chick feeds[J].Sci Food Agric,2011,(91):2378-2385.
[5]陆桂萍,曲斌,耿士伟,等.液相色谱-串联高分辨质谱测定鸡肉中卡巴氧、喹乙醇的代谢物残留[J].中国家禽,2016,38(2):33-36.Lu GP,Qu B,Geng SW,et al.Determination of metabolites of carbadox and olaquindox in chicken by LC-HRMS[J].China Poult,2016,38(2):33-36.
[6]Li E,Zhang M,Ma H,et al.Determination of3-methylquinoxaline-2-carboxylic acid of olaquindox marker residue in chicken muscles by liquid chromatography-tandem mass spectrometry[J].Chin J Chromatogr,2018,36(5):446-451.
[7]李二粉,张媚玉,马合勤,等.液相色谱-串联质谱测定鸡肉中喹乙醇残留标示物3-甲基喹恶啉-2-羧酸[J].色谱,2018,36(5):446-451.Li EF,Zhang MY,Ma HQ,et al.Determination of3-methylquinoxaline-2-carboxylic acid of olaquindox marker residue in chicken muscles by liquid chromatography-tandem mass spectrometry[J].Chin J Chromatogr,2018,36(5):446-451.
[8]欧阳姗,庞国芳,谢丽琪,等.动物组织中卡巴氧和喹乙醇以及相关代谢产物的液相色谱-串联质谱检测方法[J].分析测试学报,2008,27(6):590-594.Ou YN,Pang GF,Xie LQ,et al.Determination of the residues of carbadox,olaquindox and related metabolites in bovine and porcine muscle and liver tissues by LC-MS/MS[J].J Instrum Anal,2008,27(6):590-594.
[9]贝亦江,王扬,何丰,等.高效液相色谱法测定水产品中喹乙醇代谢物残留[J].食品科学,2013,34(10):255-258.Bei YJ,Wang Y,He F,et al.Determination of olaquindox metabolite residues in fishery products by high performance liquid chromatography[J].Food Sci,2013,34(10):255-258.
[10]赵冬艳,胡冬冬,余伟伟,等.分子印迹固相萃取-高效液相色谱联用检测饲料中喹乙醇的研究[J].粮食与饲料工业,2016,(1):72-75.Zhao DY,Hu DD,Yu WW,et al.Determination of olaquindox in feed using MISPE-HPLC[J].Cerel Feed Ind,2016,(1):72-75.
[11]Sniegocki T.Determination of carbadox and olaquindox metabolites in swine muscle by liquid chromatography/mass spectrometry[J].JChromatogr B,2014,(944):25-29.
[12]Wu Y,Yu H,Wang Y,et al.Development of a high-performance liquid chromatography method for the simultaneous quantification of quinoxaline-2-carboxylic acid and methyl-3-quinoxaline-2-carboxylic acid in animal tissues[J].J Chromatogr A,2007,1146(1):1-7.
[13]郑玲,吴玉杰,李湧,等.高效液相色谱-串联质谱法测定动物源食品中3-甲基喹喔啉-2-羧酸和喹喔啉-2-羧酸残留[J].色谱,2012,30(7):660-664.Zhen L,Wu YJ,Li Y,et al.Determination of the residues of3-methyl-quinoxaline2-carboxylic acid and quinoxaline-2-carboxylic acid in animal origin foods by high performance liquid chromatographytandem mass spectrometry[J].Chin J Chromatogr,2012,30(7):660-664.
[14]郑玲,吴玉杰,赵永锋,等.高效液相色谱-串联质谱法测定动物源食品中喹乙醇与卡巴氧残留量[J].分析测试学报,2014,33(1):21-26.Zhen L,Wu YJ,Zhao YF,et al.Determination of olaquindox and carbadox residues in animal originated food by high performance liquid chromatography-tandem mass spectrometry[J].J Instrum Anal,2014,33(1):21-26.
[15]Zhang H,Qu W,Tao Y,et al.A convenient and sensitive LC-MS/MSmethod for simultaneous determination of carbadox-and olaquindox-related residues in swine muscle and liver tissues[J].J Anal Methods Chem,2018,(2018):1-9.
[16]张静余,杨卫军,严敏鸣.液相色谱-串联质谱法测定水产品中喹乙醇和卡巴氧的代谢物残留量[J].食品安全质量检测学报,2018,9(14):3788-3793.Zhang JY,Yang WJ,Yan MM.Determination of olaquindox and carbadox metabolite residues in aquatic products by liquid chromatography-tandem mass spectrometry[J].J Food Saf Qual,2018,9(14):3788-3793.