Indirect Competitive ELISA for Detection of Amantadine,Rimantadine and Soramantadine in Chicken and Duck Meat
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摘要: 目的：建立一种检测鸡肉、鸭肉中金刚烷胺、金刚乙胺、索金刚胺的间接竞争酶联免疫吸附方法（Indirect competitive enzyme-linked immunosorbent assay，ic-ELISA）。方法：本研究基于间接竞争酶联免疫方法的原理，在酶标板微孔中预包被偶联抗原，样本中含有的金刚烷胺、金刚乙胺、索金刚胺与微孔中预包被的偶联抗原特异性地竞争酶标记抗体，催化底物显色，根据显色深浅来计算样本中金刚烷胺类药物的含量。结果：金刚烷胺、金刚乙胺、索金刚胺的检测限分别为：0.57、0.42、0.41 μg/kg （鸡肉）和0.59、0.40、0.38 μg/kg （鸭肉）；定量限分别为0.85、0.63、0.69 μg/kg （鸡肉）和0.94、0.68、0.52 μg/kg （鸭肉）；添加回收率范围为67.0%~117.9%；日内变异系数6.3%~12.7%，日间变异系数8.1%~14.5%，且实际样品检测结果与HPLC-MS一致性较高（R2=0.9990），表明该方法具有良好的准确度和精密度。结论：本研究建立的ic-ELISA方法适用于鸡鸭肉样品中金刚烷胺类药物残留的检测，方法的灵敏度高、稳定性好，可应用于批量样本的快速筛查，具有良好的实际应用价值。Abstract: Objective: To establish an ic-ELISA for the determination of amantadine,rimantadine and soramantadine in chicken and duck meat. Methods: Based on the principle of ic-ELISA,conjugated antigen was pre-coated in the wells of microtiter plate. Amantadine,rimantadine,and soramantadine contained in the samples will competitively combine with the enzyme-labelled antibody,which catalyzing colour reactions of the substrates. The concentration of amantadines in the sample was detremined on the basis of OD value. Results: The detection limits of amantadine,rimantadine and somantine were respectively 0.57,0.42,0.41 μg/kg(chicken),and 0.59,0.40,0.38 μg/kg(duck). The limits of quantification were respectively 0.85,0.63,0.69 μg/kg(chicken),and 0.94,0.68,0.52 μg/kg(duck). The added recovery range was 67.0%~117.9%;the intra-day and inter-day variation were 6.3%~12.7% and 8.1%~14.5%,respectively. The actual sample detection results were well consistent with HPLC-MS(R2=0.9990),showing that the method had good accuracy and precision. Conclusion: The ic-ELISA method established in this study is suitable for the detection of amantadine residues in chicken and duck meat samples. The method has high sensitivity and good stability. It can be used for rapid screening of batch samples and has good practical applications value.
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Keywords:
- chicken /
- duck meat /
- amantadine /
- rimantadine /
- soramantadine /
- ic-ELISA
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[1] 胡巧茹,周长朋,张玉春,等.高效液相色谱-串联质谱法测定辣椒及辣椒制品中金刚烷胺残留[J].分析测试学报,2014(10):1184-1188. [2] 赵桂宏,白向荣,李晓玲,等.金刚烷胺治疗帕金森病的有效性和安全性评价[J].中国药学杂志,2011,46(23):1847-1850. [3] 陆淳,张鑫,吴剑平,等.抗病毒药物在动物组织中残留检测方法进展[J].中国兽医杂志,2016,52(6):66-68. [4] 云环,崔凤云,严华,等.超高效液相色谱-串联质谱法测定鸡肉中的利巴韦林和金刚烷胺[J].色谱,2013,31(8):724-728. [5] FDA Prohibits.Use of antiviral drugs in poultry to help keep drugs effective for humans[EB/OL].2009-06-l8. http:Hwww.fda. Gov/NewsEvents/Newsroom/Press Announcements/2006/ucml08620.htm.
[6] 中华人民共和国农业部公告(第560号)[J].中华人民共和国农业部公报,2005,10(11):40. [7] 蒋增海,邓同炜,李文刚,等.猪群利巴韦林中毒病例[J]. 中国兽医杂志,2013,49(9):83. [8] Posyniak A,Zmudzki J,Mitrowska K. Dispersive solid-phase extraction for the determination of sulfonamides in chicken muscle by liquid chromatography[J]. Journal of Chromatography A,2005,1087(1-2):259-264.
[9] Yan H,Liu X,Cui F,et al. Determination of amantadine and rimantadine in chicken muscle by QuEChERS pretreatment method and UHPLC coupled with LTQ Orbitrap mass spectrometry[J]. Journal of Chromatography B,2013,938:8-13.
[10] You X,Yang S,Zhao J,et al. Study on the abuse of amantadine in tissues of broiler chickens by HPLC-MS/MS[J]. Journal of Veterinary Pharmacology and Therapeutics,2017,40(5):539-544.
[11] Tsuruoka Y,Nakajima T,Kanda M,et al. Simultaneous determination of amantadine,rimantadine,and memantine in processed products,chicken tissues,and eggs by liquid chromatography with tandem mass spectrometry[J]. Journal of Chromatography B,2017,1044- 1045:142-148.
[12] 邵琳智,姚仰勋,谢敏玲,等.亲水相互作用色谱-串联质谱法同时测定动物组织中金刚烷胺与利巴韦林[J].分析测试学报,2013,32(12):1448-1452. [13] Peng D,Wei W,Pan Y,et al. Preparation of a monoclonal antibody against amantadine and rimantadine and development of an indirect competitive enzyme-linked immunosorbent assay for detecting the same in chicken muscle and liver[J]. J Pharm Biomed Anal,2016,133:56-63.
[14] Wang Z,Wen K,Zhang X,et al. New hapten synthesis,antibody production,and indirect competitive enzyme-linked immnunosorbent assay for amantadine in chicken muscle[J].Food Analytical Methods,2017,11:302-308.
[15] Xu L,Shuang P,Liu L,et al. Development of sensitive and fast immunoassays for amantadine detection[J]. Food &Agricultural Immunology,2016,27(5):678-688.
[16] Wu S,Zhu F,Hu L,et al. Development of a competitive immunochromatographic assay for the sensitive detection of amantadine in chicken muscle[J]. Food Chemistry,2017,232:770-776.
[17] 吴小平,王文珺,刘萤,等.金刚烷胺ELISA试剂盒的研制[J].检验检疫学刊,2015(4):68-70. [18] 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. [19] Zhu F F,Peng J,Zhen H,et al. Specific colorimetric ELISA method based on DNA hybridization reaction and non-crosslinking gold nanoparticles aggregation for the detection of amantadine[J]. Food Chemistry,2018,257:382-387.
[20] Yu W,Zhang T,Ma M,et al. Highly sensitive visual detection of amantadine residues in poultry at the ppb level:A colorimetric immunoassay based on a Fenton reaction and gold nanoparticles aggregation[J]. Analytica Chimica Acta,2018,1027:S1451807065.
[21] 苏娟.高灵敏金刚烷胺胶体金免疫试纸条的研制[D].杭州:浙江工业大学,2015. [22] Scuracchio P S P,Poli M C C,Lemos M M M,et al. Detection of HIV-1 infection in blood donors during the immunological window period using the nucleic acid-amplification technology[J].Transfusion Medicine,2007,17(3):200-204.
[23] Barresi R. From proteins to genes:Immunoanalysis in the diagnosis of muscular dystrophies[J]. Skeletal Muscle,2011,1(1):24.
[24] Farias-Soares F L,Burrieza H P,Steiner N,et al. Immunoanalysis of dehydrins in Araucaria angustifolia embryos[J].Protoplasma,2013,250(4):911-918.
[25] Oh B R,Huang N T,Chen W,et al. Integrated nanoplasmonic sensing for cellular functional immunoanalysis using human blood[J]. ACS Nano,2014,8(3):2667-2676.
[26] Tian W,Xie H,Fu H,et al. Immunoanalysis methods for the detection of dioxins and related chemicals[J]. Sensors,2012,12(12):16710-16731.
[27] 江苏美正生物科技有限公司.一种金刚烷胺残留的快速检测试剂盒及其制备方法和应用:CN201710079790.0[P]. 2018-08-21. [28] 赵海忠,李良华,宋忠旭,等.动物抗病毒药残留检测方法研究进展[J].湖北畜牧兽医,2019,40(3):14-16. [29] 张培,陈丹.基质辅助激光解析电离飞行时间质谱快速定量有机小分子化合物应用研究[J].中国药业,2020,29(17):80-82. [30] 刘百红,熊宁,王喜亮,等.直接竞争ELISA法快速检测动物源性食品中磺胺嘧啶残留[J].中国兽药杂志,2008,42(2):16-19. [31] GB 31660.5-2019动物性食品中金刚烷胺残留量的测定-液相色谱-串联质谱法[S]. [32] GB/T 27417-2017合格评定化学分析方法确认和验证指南[S].
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