疫苗中辅料对庆大霉素残留量测定影响的考察

目的:考察用酶联免疫吸附法(ELISA)测定疫苗中庆大霉素残留量时常用辅料对测定的影响,确定ELISA方法的可行性。 方法:使用市售ELISA试剂盒,通过对配制的空白辅料溶液、疫苗样品、辅料与硫酸庆大霉素标准品混合溶液进行平行对比测定,考察疫苗中常用抑菌剂或病毒灭活剂,硫柳汞、戊二醛、甲醛和苯酚对疫苗中庆大霉素残留量测定的影响。 结果:硫柳汞、戊二醛、甲醛和苯酚本身对测定几乎没有影响,自身对照测得的硫酸庆大霉素含量为0.047~0.7 ng·L^-1,硫酸庆大霉素在4种辅料溶液中的加样回收率在77%~125%范围,2种疫苗样品中的加样回收率分别为77%和83%。 结论:ELISA方法测定疫苗中庆大霉素的残留量特异性和灵敏度均较高,且常见辅料对测定干扰不大,作为限度检查方法可行。

Objective: To determine the feasibility of the ELISA method to investigate the impact of commonly used pharmaceutical excipients on the determination of gentamycin residues in vaccine by enzyme-linked immunosorbent assay (ELISA). Methods: Using commercial ELISA kits,through the preparation of blank solution,vaccine,and the mixed solution of pharmaceutical excipients with gentamycin sulfate standard substances for parallel comparative determination,the interference of the antibacterial agents or viral inactivation agents,thiomersal,glutaraldehyde,formaldehyde and phenol on determination of gentamycin residues in vaccine was studied. Results: Thiomersal,glutaraldehyde,formaldehyde and phenol themselves have no impact on the reaction system,the gentamycin content of self-control was 0.047-0.7 ng·L^-1,the gentamycin recovery rates in 4 kinds of excipient solutions were 77%-125%,meanwhile,the gentamycin recovery rates in 2 vaccine samples were 77% and 83% respectively. Conclusion: The ELISA method has high specificity and sensitivity for determination of gentamycin residues in vaccine,and is little interfered by the common pharmaceutical excipients which is feasible as an inspection method.

1 LI Shui-ming(李水明),LIU Li(刘丽),LI Yi-jin(李逸瑾).Detection analysis of gentamincin residues in meat products(肉类食品中庆大霉素残留的检测分析).Mod Prev Med (现代预防医学),2008,5(11):2109
2 LIU Shao-gang(刘少刚),WU Jie(吴洁).The evaluation and analysis of results of the gentamicin content by ELISA(ELISA 测定疫苗中庆大霉素含量的结果评价分析).China Foreign Med Treat(中外医疗),2011,(17):128
3 YU Qi-fei(余奇飞),CHEN Cui-lian(陈翠莲),SUN Li-na(孙莉娜),et al.Development of detecting method of antibiotics residues in milk(牛奶中抗生素残留检测方法的研究进展).Acad Period Farm Prod Process(农产品加工),2005(11):72
4 Kimura R,lverson N,Sothard R.Selective lesions of the vestibular labyrinth.Ann Otol Rhinol Laryngol,1988,97:577
5 LUO Yong(罗勇).The control of antibiotic residues in vaccine development(疫苗研制中对其抗生素残留量的控制).J Pub Health Prev Med(公共卫生与预防医学),2011,22(1):88
6 ChP(中国药典).2010.Vol Ⅲ(三部):86
7 YE Xing-qian(叶兴乾),LIU Dong-hong(刘东红),CHEN Jian-chu(陈健初).Advances in the rapid detectio n of antibiotics residues in dairy products(牛奶抗生素残留快速检测技术进展及应用现状).Trans CSAE(农业工程学报),2005,21 (4):181
8 YU Ran(于然),ZHANG Rong-hua(张荣华),LIU Li-feng(刘立峰),et al.The applicability and evaluation of ELISA for detecting the residual gentamycin content in vaccine(ELISA测定疫苗中庆大霉素含量的适用性和实测结果评价).Prog Microbiol Immuol (微生物学免疫学进展),2010,38(4):29
9 YANG Xin(杨欣),ZENG Xian-wu(曾献武),LIU Yu(刘宇),et al.Kanamycin and gentamicin residues in encephalitis attenuated live vaccine were detected by ELISA method(ELISA法检测乙型脑炎减毒活疫苗中卡那霉素和庆大霉素的残留量).Prog Microbiol Immuol (微生物学免疫学进展),2010,38(3):28
10 CHEN Ling(陈玲),NIU Hua-xing(牛华星),LU Qing-quan(陆庆泉),et al.Enzyme-linked immunosorbent assay gentamicin residues in milk (酶联免疫吸附法测定牛奶中庆大霉素的残留量).Shandong J Anim Sci Vet Med(山东畜牧兽医),2009,30(11):16
11 Sherma J.Current status of pesticide residue analysis.J AOAC Int,1997,80(4):283
12 ZHAO Xiao-feng(赵晓凤),HE Fang-yang(何方洋),WAN Yu-ping(万宇平), et al.The establishment and ap plication of gentamicin ELISA detection method(庆大霉素ELISA检测方法的建立与应用).Shandong J Anim Sci Vet Med (山东畜牧兽医),2012,31(4):9
13 Yu H,Hua RH,Zhang Q,et al.Genetic evolution of swine influenza A (H3N2)viruses in China from 1970 to 2006.J Clin Microbiol,2008,46:1067