总有机碳检测法在无细胞百日咳疫苗生产清洁验证中的应用  

Application of total organic carbon method in cleaning validation during production of acellular pertussis vaccine

作　　者：谢姗姗[1] 廖丹[1] 杨勇[1] 张杜娟 杨烨[1] 赵荣丽[1] 

机构地区：[1]成都生物制品研究所有限责任公司质量检定室,四川成都610063

出　　处：《中国生物制品学杂志》2017年第11期1196-1202,1206,共8页Chinese Journal of Biologicals

摘　　要：目的证明总有机碳(total organic carbon,TOC)检测法可用于无细胞百日咳疫苗生产中的清洁验证,评价清洁方法的有效性。方法对TOC检测法的线性、精密性(重复性及中间精密性)、准确性、检测限(LOD)和定量限(LOQ)、耐用性进行验证。取不同碳浓度(20、40、80 ppm)的无细胞百日咳精制品稀释液,分别滴于玻璃、不锈钢、硅胶及塑料4种样板的表面,采用擦拭和淋洗两种方式取样,进行TOC检测,计算残留物回收率。采用TOC法对无细胞百日咳疫苗生产用具清洁效果进行评价。结果蔗糖浓度在0.25~8 mg/L范围内,与TOC值呈良好的线性关系,线性回归方程为y=1.071 7 x-0.206 3,R^2=0.999 8;重复性及中间精密性RSD值均<10%;准确性的平均回收率为99%,RSD为2.07%;超纯水的LOD及LOQ分别为24.1和80.2μg/L,棉签的LOD及LOQ分别为56.8和189.4μg/L;供试品溶液于常温下放置0、4、8、12、16、20、24、48 h后的TOC值呈逐渐降低趋势,RSD为5.66%;4种材质的淋洗和擦拭取样检测结果的RSD均<15%,回收率均在50%~120%之间。无细胞百日咳疫苗生产器具经擦拭和淋洗取样检测的残留量均≤1.5μg/cm^2。结论 TOC法可用于无细胞百日咳疫苗生产的清洁验证,证明在百日咳生产过程中所使用的清洁方法是有效的,该方法可广泛用于疫苗生产中的清洁验证。Objective To prove that the total organic carbon（TOC）method may be applied to cleaning validation during production of acellular pertussis（a P） vaccine, and evaluate the effectiveness of the cleaning methods. Methods TOC method was verified for linearity, precision（reproducibility and intermediate precision）, accuracy,limit of detection（LOD）, limit of quantitation（LOQ）and robustness. Recovery rate on the surface of different materials（e. g. glass, stainless steel, silica gel and plastic） was measured by TOC method using purified a P antigens at various carbon concentrations（20, 40 and 80 ppm） as residues. The effectiveness of cleaning methods for production apparatus of a P vaccine was evaluated by TOC method. Results The sucrose concentration at a range of 0. 25 - 8 mg/L showed good linear relationship to the TOC value, of which the linear regression equation was y = 1. 071 7 x-0. 206 3, with a R-2 value of0. 999 8. Both the RSDs in reproducibility and intermediate precision tests were less than 10%. The mean recovery rate in accuracy test was 99%, with a RSD of 2. 07%. The LOD and LOQ of ultra-pure water were 24. 1 and 80. 2 μg/L, while those of cotton bud were 56. 8 and 189. 4 μg/L, respectively. The TOC value of test samples showed a gradually decreasing tendency after storage at normal atmospheric temperature for 0, 4, 8, 12, 16, 20, 24 and 48 h, with a RSD of5. 66%. The RSDs of test samples collected by rinsing and swabbing from four kinds of materials were less than 15%,while the recovery rates were 50% - 120%. However, the residual contents of TOC in test samples collected by rinsing and swabbing were not more than 1. 5 μg/cm-2. Conclusion TOC method may be used for the cleaning validation during production of a P, which proved that the cleaning methods used for production of a P vaccine is effective and suitable for cleaning validation in vaccine production.

关 键 词：总有机碳检测法 无细胞百日咳疫苗 清洁验证 残留量 

分 类 号：R378.42[医药卫生—病原生物学] R392-33[医药卫生—基础医学]