肠道病毒71型灭活疫苗TritonX-100残留检测方法的建立与验证  

Assay development for the detection of TritonX-100 residual in inactivated enterovirus 71 vaccine

作　　者：陈晓琦[1] 王继麟[1] 郭长福[1] 明平刚[1] 张柳[1] 涂晶[1] 杨邦玲[1] 段凯[1] 黄佐林[1] 

机构地区：[1]武汉生物制品研究所有限责任公司,湖北武汉430207

出　　处：《中国病毒病杂志》2015年第2期90-95,共6页Chinese Journal of Viral Diseases

基　　金：国家"重大新药创制"科技重大专项(2013ZX09402-302);武汉市2013年高新技术成果转化及产业化计划项目(2013060803010394)

摘　　要：目的建立适合于肠道病毒71型（Vero细胞）灭活疫苗原液TritonX-100残留检测方法并加以验证。方法使用分光光度法对TritonX-100残留进行检测,并按照《中华人民共和国药典》（2010年版）二部中的相关规定对检测方法进行验证及初步应用。结果本方法在5~50μg/ml浓度范围能定量检测TritonX-100残留量,取不同浓度TritonX-100检测其回收率在85.97%~107.31%,RSD≤4.05%;重复性检测的RSD≤0.25%,中间精密度试验RSD≤3.18%;对检测中影响结果的参数：检测持续时间、检测温度、检测波长和检测试剂苯酚浓度在一定范围内适度变化后,检测理论浓度的TritonX-100,其平均回收率在86.42%~107.80%,RSD≤3.54%;340nm波长扫描PBS、5%苯酚、PBS＋苯酚的A值均〈0.01,TritonX-100及5%苯酚反应物的A值明显高于此值。结论本方法的准确度、精密度、专属性以及耐用性均符合规定,方法准确稳定,可用于对肠道病毒71型（Vero细胞）灭活疫苗原液中TritonX-100残留量的检测。Objective To develop an assay for the detection of TritonX-100 residual in inactivated enterovirus71 vaccine. Methods Spectrophotography was used to detect Triton X-100 residual in the bulk EV71 vaccine,and the validation was done according to EP（2010）in specificity,accuracy,precision and robustness.Results The linear range of this assay was 5-50μg/ml.The relative standard deviation（RSD）was below4.05%;the recovery rates were 85.97%-107.31%;the RSD detected by the same technician for 6times was below 0.25% and the RSD by two technicians at different days was below 3.18%.The recovery rates were86.42%-107.80% and RSD was blow 3.54% while parameters such time and temperature and so on vary between certain ranges.The value under 340 nm wave measured PBS,5% phenol or their mixture was below0.01 while the value of mixture of TritonX-100 and 5% phenol was over 0.01. Conclusions This established assay for detecting residual TritonX-100 showed high specificity,accuracy,precision and robustness.

关 键 词：肠道病毒71型 灭活疫苗 TritonX-100残留检测 

分 类 号：R392-33[医药卫生—免疫学]