【作者】 李荣； 艾晓辉； 邹世平； 伍刚； 徐进； 王鹏；

【Author】 LI Rong~1 AI Xiao-hui~1 ZHOU Shi-ping~1 WU Gang~1 XU Jin~1 WANG Peng~2 (1.Supervision and Testing Center for Fresh Water Fish Strain Quality,Ministry of Agriculture Yangze River Fisheries Research Institute,Jinzhou 434000,China;2.Hubei Entry-Exit Inspection and Quarantine Bureau)

【机构】 长江水产研究所农业部淡水鱼类种质监督检验测试中心； 湖北省出入境检验检疫局；

【摘要】 氯霉素(Chloramphenicol,CAP)是一种广谱抗生素,对多种病原菌有较强的抑制作用。但由于其严重的毒副作用,欧盟、美国等均规定在动物源食品中不得检出CAP,我国于2002年将氯霉素列为禁用兽药。近几年来,我国出口的鱼、虾、烤鳗等水产品和水产制品受到了严格的检测。鱼、虾肌肉组织中氯霉素残留量的检测有很多种方法,酶联免疫法(ELISA)检测水产品残留的CAP,稳定性和假阳性率以及试剂盒之间的差别未见报道。作者就两种不同厂家氯霉素试剂盒的质量和检测效果作了初步对比分析与评价,对ELISA法产生假阳性的原因进行了探讨,并运用毛细管气相色谱法(GC-ECD)和液相色谱-质谱法(LC-MS)对ELISA法中出现的阳性样品进行了验证。结果表明,用ELISA法检测鱼、虾等水产品肌肉组织中残留的氯零素时,方法简便快捷,适合大量样品的筛选。在加标回收率、检出限、重现性和假阳性率等指标上,荷兰EURO-DIAGNOSTICA氯霉素试剂盒优于德国r-Biopharm氯霉素试剂盒。两种试剂盒均对盒中所附的氯霉素标准溶液反应灵敏,重现性好,但检测鱼、虾样品,稳定性不理想,重现性较差,表明前处理后的样品仍会对酶免疫反应产生干扰。两种试剂盒测定20个鱼、虾样时检测结果不一致,相差55%。ELISA法假阳性率为10%～25%,其产生原因可能与氯霉素在鱼、虾体内的代谢物有关。氯霉素定量确认检测,液相-质谱法灵敏度高,检出限为0.1μg/kg,但仪器价格高,难以普及推广。毛细管气相色谱法具有方法简单、灵敏度较高、仪器价格较低易于推广等优点,方法检出限可达到0.1～1μg/kg。ELISA法出现的阳性样品,需用气相色谱、气相色谱-质谱和液相色谱-质谱中的任何一种方法验证。更多还原

【Abstract】 Chloramphenicol is a broad-spectrum antibiotic.It is active against a wide variety of pathogenic bacteria.But for its serious poisonous side effect,The European Union,the U.S.A.,etc.stipulate that CAP cannot be examined out from animal source food.Our country classified chloramphenicol as inhibitive animal medicine in 2002.In recent years,the aquatic products our country exported such as fish, shrimp,roast eel etc.had been strictly mensurated.There are many mensurating methods of chloramphenicol residues in fish and shrimp’s musculature.The stability,the pseudo-positive rate and the distinction of different reagent boxes have not been reported which use Enzyme-Linked Immunosorbent Assay(ELISA)to mensurate chloramphenicol residues in aquatic products.The author gave primary analysis and estimate to the quality and mensuration effect of two different reagent boxes,discussed the causes of pseudo-positive from ELISA,and used GC-ECD and LC-MS to validate the positive samples from ELISA. The result indicates that,it is simple,convenient and swift to use ELISA to mensurate chloramphenicol residues in aquatic products’ musculature such as fish and shrimp.This method is suitable for the filtration of big quantity samples.The Netherlandish EURO-DIAGNOSTICA CAP reagent boxe is better than the Germany r-Biopharm CAP reagent boxe that compared with the guidelines of reclaim rate, examine limit,recurrence and pseudo.positive.Both two reagent boxes are sensitive to the attached standard CAP.But when mensurating the fish and shrimp samples,its stability is not ideal,the recurrence is bad.This indicates that the pro-treat samples can still interfere ELISA.The result is different when we use two different reagent boxes to mensurate 20 fish and shrimp samples,55% discrepancy.The pseudo-positive rate of ELISA is 10%-25%,the reason might have something to do with the metabolite of CAP in fish and shrimp body. There is a high degree of sensitiveness when we use LC-MS for CAP fix quantify mensuration,the examine limit is 0.1μg/kg,but the price of the equipment is too high to popularize and generalize.GC-ECD has so many advantages such as simpleness,high degree of sensitiveness,low price equipment and is easy to popularize and generalize.The examine limit can reach 0.1-1μg/kg,the average reclaim rate is (78.3±6.7)%,which can satisfy the requirement of the low examine limit of CAP mensuration.When positive sample appears in ELISA,it is necessary to use GC-ECD or LC-MS to validate.更多还原