四价流感病毒裂解疫苗(MDCK细胞)中Triton X-100残留量高效液相色谱检测方法的建立及验证  

Development and verification of a method for determination of residual Triton X-100 content in quadrivalent influenza virus split vaccine(MDCK cells)by high performance liquid chromatography

作　　者：刘雪颖 张敏 何世丹 冯梦茹 任丽 罗剑 段鹏 Liu Xueying;Zhang Min;He Shidan;Feng Mengru;Ren Li;Luo Jian;Duan Peng(No.4 Research Laboratory,Shanghai Institute of Biological Products Co.,Ltd.,Shanghai 200051,China;Quality Control Department,Shanghai Institute of Biological Products Co.,Ltd.,Shanghai 201403,China)

机构地区：[1]上海生物制品研究所有限责任公司第四研究室,上海200051 [2]上海生物制品研究所有限责任公司质量检定室,上海201403

出　　处：《国际生物制品学杂志》2023年第4期204-208,共5页International Journal of Biologicals

基　　金：上海市科委科技攻关计划(21S11903200)。

摘　　要：目的建立并验证四价流感病毒裂解疫苗(MDCK细胞)中Triton X-100残留量高效液相色谱(high performance liquid chromatography,HPLC)检测方法。方法应用HPLC法检测四价流感病毒裂解疫苗中Triton X-100残留量,并对该方法的系统适用性、专属性、线性、准确度、精密度、耐用性、定量限及检测限进行验证。结果HPLC法检测Triton X-100的理论塔板数在1560~1599之间,峰面积和保留时间的相对标准偏差(relative standard deviation,RSD)分别为1.93%和0.30%,且Triton X-100色谱峰与杂质分离良好;空白溶液无干扰峰出现;Triton X-100的浓度在5~160μg/ml范围内时,浓度与峰面积呈良好的线性关系,回归曲线决定系数为0.999;检测Triton X-100样品的加标回收率均在95%~105%之间,RSD均不超过2%;同一实验人员重复进样6次检测峰面积的RSD为0.58%,同一实验人员在不同工作日和不同检测人员在同一工作日检测峰面积的RSD也均小于2%;改变柱温后,峰面积检测值的RSD为0.45%;定量限为5μg/ml,检测限为1μg/ml。结论成功建立了四价流感病毒裂解疫苗中Triton X-100残留量HPLC检测方法,该方法的系统适用性、专属性、线性、准确度、精密度、耐用性良好。ObjectiveTo develop and verify a high performance liquid chromatography(HPLC)method for determination of residual Triton X-100 content in quadrivalent influenza virus split vaccine(MDCK cells).MethodsTriton X-100 residue in the vaccine was detected by HPLC,and the system applicability,specificity,linearity,accuracy,precision,durability,quantification limit,and detection limit of this method were verified.ResultsThe numbers of theoretical plates of Triton X-100 detected by HPLC were 1560-1599.The relative standard deviations(RSDs)of peak area and retention time of Triton X-100 were 1.93%and 0.30%,respectively.The Triton X-100 chromatographic peak and impurity separation was good.No interference peak appeared in the blank solution.In the range of 5-160μg/ml,the Triton X-100 concentration showed good linear relationship to the peak area(coefficient of determination at 0.999).The spike recovery rates of Triton X-100 at various concentrations were 95%-105%,all with RSDs no more than 2%.The RSDs of peak area of 6 samples tested by one staff was 0.58%,and the RSDs of peak area by one staff on various working days and by various staff on one working day were all less than 2%.After the change of column temperature,the RSD of peak area was 0.45%.The limits of detection and quantitation were 1 and 5μg/ml,respectively.ConclusionsHPLC detection method for Triton X-100 residue in quadrivalent influenza virus split vaccine is successfully established.The system applicability,specificity,linearity,accuracy,precision,and durability of this method are good.

关 键 词：流感疫苗 Madin Darby犬肾细胞 色谱法 高效液相 曲拉通X-100 

分 类 号：R392-3[医药卫生—免疫学]