液相色谱-串联质谱法快速测定鸡蛋中金刚烷胺、氟苯尼考和氟苯尼考胺残留量
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摘要
目的建立同时测定鸡蛋中金刚烷胺、氟苯尼考及氟苯尼考代谢物氟苯尼考胺残留的液相色谱-串联质谱(liquid chromatography-tandem mass spectrometry, LC-MS/MS)快速分析方法。方法鸡蛋样品用1.0%乙酸乙腈提取,提取后用PSA和C_(18)净化,氮吹后用流动相复溶,复溶液过滤膜后进行液相色谱-串联质谱测定。分析采用XBridge C_(18)色谱柱,以0.02%甲酸溶液和甲醇乙腈混合溶剂为流动相进行梯度洗脱,正负离子切换监测模式下进行监测,同位素内标法定量。结果3种化合物在一定浓度(氟苯尼考为0.10～20μg/L,金刚烷胺和氟苯尼考胺为0.5～100μg/L)范围内具有较好的线性关系,金刚烷胺、氟苯尼考和氟苯尼考胺的检出限(limit of detection, LOD)分别为0.3、0.06、0.3μg/kg,定量限(limit of quantitation, LOQ)分别为1.0、0.20、1.0μg/kg。当3种化合物在鸡蛋中的加标水平为LOQ、5LOQ和10LOQ时,平均回收率在94.9%～102%之间,相对标准偏差范围为1.2%～4.9%。结论方法简便、快速、准确,能满足鸡蛋中金刚烷胺和氟苯尼考残留量同时分析需要。
Objective To establish a rapid method for simultaneous determination of amantadine, florfenicol and florfenicol amine residues in eggs by liquid chromatography-tandem mass spectrometry(LC-MS/MS). Methods Egg samples were extracted with 1.0% acetonitrile acetate, purified with PSA and C_(18), and dissolved with mobile phase after nitrogen blowing, detected by LC-MS/MS after filtering the solution. An XBridge C_(18) column was used for the separation by gradient elution using a mixture of 0.02% formic acid solution and methanol/acetonitrile as the mobile phase, and a mass spectrometer was used in the positive and negative ion mode. The samples were quantified with the isotope dilution and internal standard methods. Results Three compounds had good linear relationships in the range of 0.10-20 μg/L(florfenicol) or 0.50-100 μg/L(amantadine and florfenicol amine). The limits of detection of amantadine, florfenicol and florfenicol amine were 0.30, 0.06, and 0.30 μg/kg, respectively. The limits of quantitation of amantadine, florfenicol and florfenicol amine were 1.0, 0.20, and 1.0 μg/kg, respectively. The recoveries at 3 spiked levels(LOQ, 5 LOQ, 10 LOQ) were 94.9%-102%, with the relative standard deviations of1.2%-4.9%. Conclusion This method is simple, rapid and accurate, which is suitable for simultaneous determination of amantadine, florfenicol and florfenicol residues in eggs.
Objective To establish a rapid method for simultaneous determination of amantadine, florfenicol and florfenicol amine residues in eggs by liquid chromatography-tandem mass spectrometry(LC-MS/MS). Methods Egg samples were extracted with 1.0% acetonitrile acetate, purified with PSA and C_(18), and dissolved with mobile phase after nitrogen blowing, detected by LC-MS/MS after filtering the solution. An XBridge C_(18) column was used for the separation by gradient elution using a mixture of 0.02% formic acid solution and methanol/acetonitrile as the mobile phase, and a mass spectrometer was used in the positive and negative ion mode. The samples were quantified with the isotope dilution and internal standard methods. Results Three compounds had good linear relationships in the range of 0.10-20 μg/L(florfenicol) or 0.50-100 μg/L(amantadine and florfenicol amine). The limits of detection of amantadine, florfenicol and florfenicol amine were 0.30, 0.06, and 0.30 μg/kg, respectively. The limits of quantitation of amantadine, florfenicol and florfenicol amine were 1.0, 0.20, and 1.0 μg/kg, respectively. The recoveries at 3 spiked levels(LOQ, 5 LOQ, 10 LOQ) were 94.9%-102%, with the relative standard deviations of1.2%-4.9%. Conclusion This method is simple, rapid and accurate, which is suitable for simultaneous determination of amantadine, florfenicol and florfenicol residues in eggs.
引文
[1]中华人民共和国农业部.关于清查金刚烷胺等抗病毒药物的紧急通知[Z]. 2015-12-02.Ministry of Agriculture of the People's Republic of China. Urgent notice for checking the antiviral drugs such as amantadine[Z]. 2015-12-02.
[2]中华人民共和国农业部.中华人民共和国农业部第235号公告[Z].2002-12-24.Ministry of Agriculture of the People's Republic of China. The Ministry of Agriculture Bulletin of PRC235[Z]. 2002-12-24.
[3]刘智宏,黄耀凌,汪霞,等.水产品中甲砜霉素、氟苯尼考和氟苯尼考胺酶联免疫多残留测定[J].中国兽药杂志,2010,44(12):1-5.Liu ZH, Huang YL, Wang X, et al. Multi-residues determination of thiamphenicol, florfenicol, florfenicol amine in aquatic products by ELISA[J]. Chin J Vet Drug, 2010,44(12):1-5.
[4]冯才伟,黄健,王兆芹,等.检测动物组织中金刚烷胺残留的酶联免疫试剂盒的研制[J].畜牧与兽医,2016,48(11):27-31.Feng C, Huang J, Wang Z, et al. Development of an enzyme-linked immunosorbent assay kit for the determination of the amantadine residues in animal tissues[J]. Anim Husb Vet Med, 2016,48(11):27-31.
[5]胡红美,郭远明,雷科,等.分散固相萃取净化-气相色谱法测定水产品中氯霉素和氟苯尼考[J].食品科学,2014,(8):231-235.Hu H, Guo Y, Lei K, et al. Determination of chloramphenicol and florfenicol in fishery products by using dispersive solid phase extraction and gas chromatography[J]. Food Sci, 2014,(8):231-235.
[6]刘永涛,艾晓辉,李荣,等.水产品中氟苯尼考和氟苯尼考胺的高效液相色谱荧光法同时测定[J].分析测试学报,2008, 27(3):316-318.Liu YT, Ai XH, Li R, et al. Simultaneous determination of florfenicol and florfenicol amine in aquatic products by HPLC-FD[J]. J Instrum Anal,2008, 27(3):316-318.
[7] Xie K, Jia L, Yao Y, et al. Simultaneous determination of thiamphenicol,florfenicol and florfenicol amine in eggs by reversed-phase high-performance liquid chromatography with fluorescence detection[J]. J Chromatogr B, 2011,(879):2351-2354.
[8]吴银良,赵健,叶宇飞,等.同位素稀释液相色谱-串联质谱法测定鸡蛋中金刚烷胺类药物残留量[J].分析测试学报,2014,33(8):905-910.Wu YL, Zhao J, Ye YF, et al. Simultaneous determination of amantadine,rimantadine and memantine residues in eggs by liquid chromatography with tandem mass spectrometry[J]. J Instrum Anal, 2014, 33(8):905-910.
[9]路川,白翔,桑鲁燕,等.分子印迹-液相色谱-串联质谱法检测鸡肉中金刚烷胺残留[J].食品安全质量检测学报,2017, 8(11):4169-4173.Lu C, Bai X, Sang LY, et al. Determination of amantadine residues in chicken by molecularly imprinted-liquid chromatography-tandem mass spectrometry[J]. J Food Saf Qual, 2017, 8(11):4169-4173.
[10]段科,刘刚,娄喜山,等.超高效液相色谱-串联质谱法测定动物组织中的金刚烷胺、金刚乙胺和盐酸美金刚[J].食品安全质量检测学报,2018,9(3):652-658.Duan K, Liu G, Lou XS, et al. Determination of am antadine, rimantadine and memantine hydrochloride in animal tissue by ultra performance liquid chromatography-tandem mass spectrometry[J]. J Food Saf Qual, 2018,9(3):652-658.
[11]尹晖,孙雷,毕言锋,等.鸡肉和鸡蛋中金刚烷胺与金刚乙胺残留检测UPLC-MS/MS法研究[J].中国兽医杂志,2014,48(6):32-35.Yin H, Sun L, Bi YF, et al. Determination of amantadine and rimantadine residue in chicken muscle and egg by UPLC-MS/MS[J]. Chin J Vet Drug,2014,48(6):32-35.
[12]林涛,樊建麟,刘兴勇,等.分散固相萃取净化-超高效液相色谱-串联质谱法测定鸡蛋和鸡肉中金刚烷胺和金刚乙胺残留[J].色谱,2015,33(11):1169-1174.Lin T, Fan J, Liu X, et al. Determination of amantadine and rimantadine residues in egg and chicken samples by dispersive solid phase extraction purification-ultra high performance liquid chromatography tandem mass spectrometry[J]. Chin J Chromatogr, 2015, 33(11):1169-1174.
[13] Suo D, Wang P, Li Y, et al. Simultaneous determination of antibiotics and amantadines in animal-derived feedstuffs by ultraperformance liquid chromatographic-tandem mass spectrometry[J]. J Chromatogr B, 1095:183-190.
[14]曲斌,朱志谦,陆桂萍,等.UPLC-MS/MS快速测定鸡肉中金刚烷胺和氟喹诺酮类药物残留[J].中国兽药杂志,2013,47(7):50-54.Qu B, Zhu Z, Lu G, et al. Rapid determination of amantadine and fluoroquinolone residues in poultry muscle by UPLC-MS/MS method[J].Chin J Vet Drug, 2013,47(7):50-54.
[15]王威利,吴维烽,何绮霞,等.高效液相色谱-串联质谱法同时检测鸡蛋中金刚烷胺与四种氟喹诺酮类药物残留的研究[J].中国家禽,2016,38(2):29-32.Wang W, Wu W, He QX, et al. Simultaneous determination of amantadine and four kinds of quinolones in eggs by high performance liquid chromatography with tandem mass spectrometry[J]. China Poul, 2016,38(2):29-32.
[16]曾勇,董文婷,龚波,等.超高效液相色谱-串联质谱法快速测定禽蛋中氯霉素、氟苯尼考及其代谢物残留[J].中国兽药杂志,2017,51(12):42-45.Zeng Y, Dong W, Gong B, et al. Rapid determination of chloramphenicol,florfenicol and its metabolites in poultry eggs by UPLC-MS/MS[J].Chin J Vet Drug, 2017, 51(12):42-45.
[17]张静,高玉时,周倩,等.高效液相色谱-电喷雾电离串联质谱法测定鸡肉、鸡蛋中氯霉素、甲砜霉素和氟苯尼考药物残留量[J].食品安全质量检测学报,2017, 8(6):2211-2219.Zhang J, Gao YS, Zhou Q, et al. Determination of chlorampheincol,thiamphenicol and florfenicol residues in chicken and egg by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry[J]. J Food Saf Qual, 2017, 8(6):2211-2219.
[18]张凤清,王岩松,范世华,等.高效液相色谱-串联质谱法测定动物肌肉组织中氯霉素、甲砜霉素和氟苯尼考残留量[J].食品科学,2010,31(8):248-251.Zhang F, Wang Y, Fan S, et al. High performance liquid chromatography-tandem mass spectrometric determination of chlorampheincol, thiamphenicol and florfenicol residues in animal muscles[J]. Food Sci, 2010,31(8):248-251.
[2]中华人民共和国农业部.中华人民共和国农业部第235号公告[Z].2002-12-24.Ministry of Agriculture of the People's Republic of China. The Ministry of Agriculture Bulletin of PRC235[Z]. 2002-12-24.
[3]刘智宏,黄耀凌,汪霞,等.水产品中甲砜霉素、氟苯尼考和氟苯尼考胺酶联免疫多残留测定[J].中国兽药杂志,2010,44(12):1-5.Liu ZH, Huang YL, Wang X, et al. Multi-residues determination of thiamphenicol, florfenicol, florfenicol amine in aquatic products by ELISA[J]. Chin J Vet Drug, 2010,44(12):1-5.
[4]冯才伟,黄健,王兆芹,等.检测动物组织中金刚烷胺残留的酶联免疫试剂盒的研制[J].畜牧与兽医,2016,48(11):27-31.Feng C, Huang J, Wang Z, et al. Development of an enzyme-linked immunosorbent assay kit for the determination of the amantadine residues in animal tissues[J]. Anim Husb Vet Med, 2016,48(11):27-31.
[5]胡红美,郭远明,雷科,等.分散固相萃取净化-气相色谱法测定水产品中氯霉素和氟苯尼考[J].食品科学,2014,(8):231-235.Hu H, Guo Y, Lei K, et al. Determination of chloramphenicol and florfenicol in fishery products by using dispersive solid phase extraction and gas chromatography[J]. Food Sci, 2014,(8):231-235.
[6]刘永涛,艾晓辉,李荣,等.水产品中氟苯尼考和氟苯尼考胺的高效液相色谱荧光法同时测定[J].分析测试学报,2008, 27(3):316-318.Liu YT, Ai XH, Li R, et al. Simultaneous determination of florfenicol and florfenicol amine in aquatic products by HPLC-FD[J]. J Instrum Anal,2008, 27(3):316-318.
[7] Xie K, Jia L, Yao Y, et al. Simultaneous determination of thiamphenicol,florfenicol and florfenicol amine in eggs by reversed-phase high-performance liquid chromatography with fluorescence detection[J]. J Chromatogr B, 2011,(879):2351-2354.
[8]吴银良,赵健,叶宇飞,等.同位素稀释液相色谱-串联质谱法测定鸡蛋中金刚烷胺类药物残留量[J].分析测试学报,2014,33(8):905-910.Wu YL, Zhao J, Ye YF, et al. Simultaneous determination of amantadine,rimantadine and memantine residues in eggs by liquid chromatography with tandem mass spectrometry[J]. J Instrum Anal, 2014, 33(8):905-910.
[9]路川,白翔,桑鲁燕,等.分子印迹-液相色谱-串联质谱法检测鸡肉中金刚烷胺残留[J].食品安全质量检测学报,2017, 8(11):4169-4173.Lu C, Bai X, Sang LY, et al. Determination of amantadine residues in chicken by molecularly imprinted-liquid chromatography-tandem mass spectrometry[J]. J Food Saf Qual, 2017, 8(11):4169-4173.
[10]段科,刘刚,娄喜山,等.超高效液相色谱-串联质谱法测定动物组织中的金刚烷胺、金刚乙胺和盐酸美金刚[J].食品安全质量检测学报,2018,9(3):652-658.Duan K, Liu G, Lou XS, et al. Determination of am antadine, rimantadine and memantine hydrochloride in animal tissue by ultra performance liquid chromatography-tandem mass spectrometry[J]. J Food Saf Qual, 2018,9(3):652-658.
[11]尹晖,孙雷,毕言锋,等.鸡肉和鸡蛋中金刚烷胺与金刚乙胺残留检测UPLC-MS/MS法研究[J].中国兽医杂志,2014,48(6):32-35.Yin H, Sun L, Bi YF, et al. Determination of amantadine and rimantadine residue in chicken muscle and egg by UPLC-MS/MS[J]. Chin J Vet Drug,2014,48(6):32-35.
[12]林涛,樊建麟,刘兴勇,等.分散固相萃取净化-超高效液相色谱-串联质谱法测定鸡蛋和鸡肉中金刚烷胺和金刚乙胺残留[J].色谱,2015,33(11):1169-1174.Lin T, Fan J, Liu X, et al. Determination of amantadine and rimantadine residues in egg and chicken samples by dispersive solid phase extraction purification-ultra high performance liquid chromatography tandem mass spectrometry[J]. Chin J Chromatogr, 2015, 33(11):1169-1174.
[13] Suo D, Wang P, Li Y, et al. Simultaneous determination of antibiotics and amantadines in animal-derived feedstuffs by ultraperformance liquid chromatographic-tandem mass spectrometry[J]. J Chromatogr B, 1095:183-190.
[14]曲斌,朱志谦,陆桂萍,等.UPLC-MS/MS快速测定鸡肉中金刚烷胺和氟喹诺酮类药物残留[J].中国兽药杂志,2013,47(7):50-54.Qu B, Zhu Z, Lu G, et al. Rapid determination of amantadine and fluoroquinolone residues in poultry muscle by UPLC-MS/MS method[J].Chin J Vet Drug, 2013,47(7):50-54.
[15]王威利,吴维烽,何绮霞,等.高效液相色谱-串联质谱法同时检测鸡蛋中金刚烷胺与四种氟喹诺酮类药物残留的研究[J].中国家禽,2016,38(2):29-32.Wang W, Wu W, He QX, et al. Simultaneous determination of amantadine and four kinds of quinolones in eggs by high performance liquid chromatography with tandem mass spectrometry[J]. China Poul, 2016,38(2):29-32.
[16]曾勇,董文婷,龚波,等.超高效液相色谱-串联质谱法快速测定禽蛋中氯霉素、氟苯尼考及其代谢物残留[J].中国兽药杂志,2017,51(12):42-45.Zeng Y, Dong W, Gong B, et al. Rapid determination of chloramphenicol,florfenicol and its metabolites in poultry eggs by UPLC-MS/MS[J].Chin J Vet Drug, 2017, 51(12):42-45.
[17]张静,高玉时,周倩,等.高效液相色谱-电喷雾电离串联质谱法测定鸡肉、鸡蛋中氯霉素、甲砜霉素和氟苯尼考药物残留量[J].食品安全质量检测学报,2017, 8(6):2211-2219.Zhang J, Gao YS, Zhou Q, et al. Determination of chlorampheincol,thiamphenicol and florfenicol residues in chicken and egg by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry[J]. J Food Saf Qual, 2017, 8(6):2211-2219.
[18]张凤清,王岩松,范世华,等.高效液相色谱-串联质谱法测定动物肌肉组织中氯霉素、甲砜霉素和氟苯尼考残留量[J].食品科学,2010,31(8):248-251.Zhang F, Wang Y, Fan S, et al. High performance liquid chromatography-tandem mass spectrometric determination of chlorampheincol, thiamphenicol and florfenicol residues in animal muscles[J]. Food Sci, 2010,31(8):248-251.