化学发光免疫试剂盒检测猪肉中呋喃西林代谢物的残留
|
摘要
为了快速检测猪肉中呋喃西林代谢物残留量,试验采用自主研发的以磁性微球为固定相的化学发光免疫试剂盒,并对其方法学进行评价及与高效液相色谱-串联质谱法(HPLC-MS/MS法)检测结果进行比较分析。结果表明:采用化学发光免疫试剂盒测定猪肉中呋喃西林代谢物的灵敏度为0.331μg/kg,精密度较好,最低检测限为0.077μg/kg,与呋喃西林原药的交叉反应率为8.9%,与其他类似物的交叉反应率均<0.1%,平均回收率为92.7%~104.8%,与HPLC-MS/MS法检测样本的阴性和阳性结果一致。说明该试剂盒具有灵敏度高、特异性强、检测速度快等特点,适用于现场快速检测猪肉中呋喃西林代谢物的残留。
The aim of the present study was to quickly detect nitrofurazone metabolite residues in pork. A chemiluminescent enzyme immunoassay kit based on magnetic beads was utilized to detecting semicarbazide residues in pork quantitative. The methodology was evaluated and chemiluminescent enzyme immunoassay kit was also compared with HPLC-MS/MS on the spot. The results showed that the sensitivity of determination of itrofurazone metabolites in pork by magnetic beads chemiluminescence methodwas 0. 331 μg/kg.The precision was good and the limit of detection was 0. 077 μg/kg. The cross-reactivity value with nitrofurazone was 8. 9%,and with other analogues was less than 0. 1%. The average recovery of negative samples ranged from 92. 7% to 104. 8%. The results indicated that the kit had the advantages of high sensitivity,strong specificity and high detection speed. It was suitable for rapid detection of nitrofurazone metabolite residues in pork on the spot.
The aim of the present study was to quickly detect nitrofurazone metabolite residues in pork. A chemiluminescent enzyme immunoassay kit based on magnetic beads was utilized to detecting semicarbazide residues in pork quantitative. The methodology was evaluated and chemiluminescent enzyme immunoassay kit was also compared with HPLC-MS/MS on the spot. The results showed that the sensitivity of determination of itrofurazone metabolites in pork by magnetic beads chemiluminescence methodwas 0. 331 μg/kg.The precision was good and the limit of detection was 0. 077 μg/kg. The cross-reactivity value with nitrofurazone was 8. 9%,and with other analogues was less than 0. 1%. The average recovery of negative samples ranged from 92. 7% to 104. 8%. The results indicated that the kit had the advantages of high sensitivity,strong specificity and high detection speed. It was suitable for rapid detection of nitrofurazone metabolite residues in pork on the spot.
引文
[1]刘书贵,吴仕辉,郑光明,等.呋喃西林代谢物在杂交鳢(斑鳢♀×乌鳢♂)体内的残留消除规律研究[J].华南农业大学学报,2013,34(2):248-253.
[2]李东利,李健,常志强,等.呋喃西林代谢产物在中国对虾(Fenneropenaeus chinensis)体内的消除规律及其对代谢酶活性的影响[J].渔业科学进展,2015,36(5):87-94.
[3]DU N N,CHEN M M,SHENG L Q,et al.Determination of nitrofuran metabolites in shrimp by high performance liquid chromatography with fluorescence detection and liquid chromatography-tandem mass spectrometry using a new derivatization reagent[J].J Chromatogr A,2014,1327:90-96.
[4]闫叶娜.呋喃西林代谢物单克隆抗体的制备及其免疫分析方法的建立[D].南京:南京农业大学,2010.
[5]MCCRACKEN R J,KENNEDY D G.Detection,accumulation and distribution of nitrofuran residues in egg yolk,albumen and shell[J].Food Addit Contam,2007,24(1):26-33.
[6]CHU P S,LOPEZ M I,ABRAHAM A,et al.Residue depletion of nitrofuran drugs and their tissue-bound metabolites in channel catfish(Ictalurus punctatus)after oral dosing[J].J Agric Food Chem,2008,56(17):8030-8034.
[7]何进,谢体波,何方洋.呋喃西林代谢物直接竞争CMIA检测法的研究[J].食品与发酵科技,2016,52(3):87-92.
[8]佘永新,李宝海,曹维强,等.动物性食品中硝基呋喃类药物及其代谢物残留检测技术的研究进展[J].现代科学仪器,2010(6):149-153.
[9]李芳,陈莹,李献刚,等.动物源性食品中硝基呋喃及其代谢物产物检测方法研究进展[J].食品安全质量检测学报,2016,7(6):2320-2327.
[10]魏法山,盖圣美,谢文佳,等.动物源性食品中硝基呋喃类兽药残留检测方法的研究进展[J].食品安全质量检测学报,2016,7(6):2289-2295.
[11]VERDON E,COUEDOR P,SANDERS P.Multi-residue monitoring for the simultaneous determination of five nitrofurans(furazolidone,furaltadone,nitrofurazone,nitrofurantoine,nifursol)in poultry muscle tissue through the detection of their five major metabolites(AOZ,AMOZ,SEM,AHD,DNSAH)by liquid chromatography coupled to electrospray tandem mass spectrometry-in-house validation in line with Commission Decision 657/2002/EC[J].Anal Chim Acta,2007,586(1/2):336-347.
[12]吴鹏,万宇平,陶光灿,等.呋喃西林代谢物酶联免疫检测方法的建立[J].安徽农业科学,2013,41(5):1969-1971.
[13]黄登宇,高丽霞,李亚楠,等.间接竞争化学发光酶免疫法检测动物源食品中的呋喃西林代谢物[J].食品安全质量检测学报,2017,8(2):402-410.
[14]彭宏威,白瑞樱,陈殿,等.响应面试验优化动物源性食品中呋喃西林代谢物ELISA检测的衍生条件[J].食品科学,2017,38(6):287-294.
[15]国家质量监督检验检疫总局,国家标准化管理委员会.GB/T21311—2007动物源性食品中硝基呋喃类药物代谢物残留量检测方法:高效液相色谱/串联质谱法[S].北京:中国标准出版社,2007.
[16]MOTTIER P,KHONG S P,GREMAUD E,et al.Quantitative determination of four nitrofuran metabolites in meat by isotope dilution liquid chromatography-electrospray ionisation-tandem mass spectrometry[J].J Chromatogr A,2005,1067(1/2):85-91.
[17]刘柏林,谢继安,赵紫微,等.超高效液相色谱-串联质谱法测定动物源性食品中硝基呋喃代谢物[J].食品安全质量检测学报,2014,5(2):421-427.
[18]国家质量监督检验检疫总局,国家标准化管理委员会.GB/T20752—2006猪肉、牛肉、鸡肉、猪肝和水产品中硝基呋喃类代谢物残留量的测定:液相色谱-联质谱法[S].北京:中国标准出版社,2006.
[19]LEE H S,LEE H E,PARK D J,et al.Clinical significance of serum and tissue Dickkopf-1 levels in patients with gastric cancer[J].Clin Chim Acta,2012,413(21/22):1753-1760.
[2]李东利,李健,常志强,等.呋喃西林代谢产物在中国对虾(Fenneropenaeus chinensis)体内的消除规律及其对代谢酶活性的影响[J].渔业科学进展,2015,36(5):87-94.
[3]DU N N,CHEN M M,SHENG L Q,et al.Determination of nitrofuran metabolites in shrimp by high performance liquid chromatography with fluorescence detection and liquid chromatography-tandem mass spectrometry using a new derivatization reagent[J].J Chromatogr A,2014,1327:90-96.
[4]闫叶娜.呋喃西林代谢物单克隆抗体的制备及其免疫分析方法的建立[D].南京:南京农业大学,2010.
[5]MCCRACKEN R J,KENNEDY D G.Detection,accumulation and distribution of nitrofuran residues in egg yolk,albumen and shell[J].Food Addit Contam,2007,24(1):26-33.
[6]CHU P S,LOPEZ M I,ABRAHAM A,et al.Residue depletion of nitrofuran drugs and their tissue-bound metabolites in channel catfish(Ictalurus punctatus)after oral dosing[J].J Agric Food Chem,2008,56(17):8030-8034.
[7]何进,谢体波,何方洋.呋喃西林代谢物直接竞争CMIA检测法的研究[J].食品与发酵科技,2016,52(3):87-92.
[8]佘永新,李宝海,曹维强,等.动物性食品中硝基呋喃类药物及其代谢物残留检测技术的研究进展[J].现代科学仪器,2010(6):149-153.
[9]李芳,陈莹,李献刚,等.动物源性食品中硝基呋喃及其代谢物产物检测方法研究进展[J].食品安全质量检测学报,2016,7(6):2320-2327.
[10]魏法山,盖圣美,谢文佳,等.动物源性食品中硝基呋喃类兽药残留检测方法的研究进展[J].食品安全质量检测学报,2016,7(6):2289-2295.
[11]VERDON E,COUEDOR P,SANDERS P.Multi-residue monitoring for the simultaneous determination of five nitrofurans(furazolidone,furaltadone,nitrofurazone,nitrofurantoine,nifursol)in poultry muscle tissue through the detection of their five major metabolites(AOZ,AMOZ,SEM,AHD,DNSAH)by liquid chromatography coupled to electrospray tandem mass spectrometry-in-house validation in line with Commission Decision 657/2002/EC[J].Anal Chim Acta,2007,586(1/2):336-347.
[12]吴鹏,万宇平,陶光灿,等.呋喃西林代谢物酶联免疫检测方法的建立[J].安徽农业科学,2013,41(5):1969-1971.
[13]黄登宇,高丽霞,李亚楠,等.间接竞争化学发光酶免疫法检测动物源食品中的呋喃西林代谢物[J].食品安全质量检测学报,2017,8(2):402-410.
[14]彭宏威,白瑞樱,陈殿,等.响应面试验优化动物源性食品中呋喃西林代谢物ELISA检测的衍生条件[J].食品科学,2017,38(6):287-294.
[15]国家质量监督检验检疫总局,国家标准化管理委员会.GB/T21311—2007动物源性食品中硝基呋喃类药物代谢物残留量检测方法:高效液相色谱/串联质谱法[S].北京:中国标准出版社,2007.
[16]MOTTIER P,KHONG S P,GREMAUD E,et al.Quantitative determination of four nitrofuran metabolites in meat by isotope dilution liquid chromatography-electrospray ionisation-tandem mass spectrometry[J].J Chromatogr A,2005,1067(1/2):85-91.
[17]刘柏林,谢继安,赵紫微,等.超高效液相色谱-串联质谱法测定动物源性食品中硝基呋喃代谢物[J].食品安全质量检测学报,2014,5(2):421-427.
[18]国家质量监督检验检疫总局,国家标准化管理委员会.GB/T20752—2006猪肉、牛肉、鸡肉、猪肝和水产品中硝基呋喃类代谢物残留量的测定:液相色谱-联质谱法[S].北京:中国标准出版社,2006.
[19]LEE H S,LEE H E,PARK D J,et al.Clinical significance of serum and tissue Dickkopf-1 levels in patients with gastric cancer[J].Clin Chim Acta,2012,413(21/22):1753-1760.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |