基于连续流动注射安培法定量检测多糖蛋白结合疫苗中氰化物的残留量  

Quantitative determination of cyanide residues in polysaccharide-protein conjugate vaccines based on flow injection analysis(FIA) with amperometric detector

作　　者：张姮婕 唐瑶 蒋佳兴 马晶 李炎 王叔桥 杨蕾 ZHANG Hengjie;TANG Yao;JIANG Jiaxing;MA Jing;LI Yan;WANG Shuqiao;YANG Lei(Sichuan Institute for Drug Control,NMPA Key Laboratory for Quality Control and Evaluation of Vaccines and Biological Products,SCMPA Key Laboratory for Quality Monitoring and Risk Assessment of Biological Products,Chengdu,611731,Sichuan Province,China)

机构地区：[1]四川省药品检验研究院,国家药品监督管理局疫苗及生物制品质量控制与评价重点实验室,四川省药品监督管理局生物制品质量监测与风险评估重点实验室,四川成都611731

出　　处：《中国生物制品学杂志》2023年第3期352-356,362,共6页Chinese Journal of Biologicals

基　　金：四川省自然科学基金(2022NSFSC1572);国家药典委员会药品标准提高课题(2021S08)。

摘　　要：目的 建立基于连续流动注射安培法的多糖蛋白结合疫苗中氰化物残留量的检测方法,并进行验证。方法 超滤法去除样品中的大分子物质后,采用3700全自动化学分析仪检测氰化物残留量,进样时间为:35 s;进样体积为:200μL;泵速为:40%;样品周期时间为:140 s;紫外波长为:312 nm;安培检测器。验证方法的专属性、基质效应、线性范围、检出限、定量限、准确性、精密性及稳定性。采用建立的方法检测5个厂家生产的b型流感嗜血杆菌结合疫苗及A群C群脑膜炎球菌结合疫苗多糖衍生物原液(13批)、结合物原液(21批)中氰化物残留量。结果 空白样品对检测无干扰;b型流感嗜血杆菌结合疫苗多糖衍生物和结合物原液、A群C群脑膜炎球菌结合疫苗多糖衍生物和结合物原液基质效应溶液的回收率分别为97.4%、102.4%、96.8%、99.8%,CV均<15%;氰基浓度在0.312 5~80 ng/mL范围内,与峰高呈良好的线性关系,回归方程为:y=133.13 x+57.556,R^(2)=0.999 1;检出限为0.2 ng/mL,定量限为0.6 ng/mL;加入氰基浓度为5、10、20 ng/mL对照溶液样品的回收率均值分别为108.9%、106.5%、103.5%,RSD为6.4%;精密性验证的CV均<15%;氰基浓度为80、40、20、5 ng/mL的对照溶液连续进样20针的平均浓度分别为76、38、18、5 ng/mL,CV均<15%。13批多糖衍生物原液样品均有检出氰化物残留,21批结合物原液样品中2批未检出,其他均有检出。结论 本研究建立的方法具有良好的准确性、精密性及稳定性,可应用于多糖蛋白结合疫苗中氰化物残留量的定量检测。Objective To develop and verify a method for determination of cyanide residues in polysaccharide-protein conjugate vaccines based on flow injection analysis(FIA)with amperometric detector.Methods After removing macromolecular substances in the samples by ultrafiltration,the cyanide residues were determined by a 3700 automatic chemical analyzer with the injection time of 35 s,injection volume of 200 μL,pump speed of 40%,sample cycle time of 140 s,ultraviolet wavelength of 312 nm and an Ampere detector.The developed method was verified for the specificity,matrix effect,linear range,limit of detection(LOD),limit of quantitation(LOQ),accuracy,presicion and stability.The cyanide residues in the polysaccharide derivative bulk(13 batches)and conjugate bulk(21 batches)of Haemophilus influenzae type b conjugate vaccine and group A and C meningococcal conjugate vaccine produced by 5 manufacturers were determined by the developed method.Results Blank samples showed no interference to detection;The recoveries of the matrix effector solution of Haemophilus influenzae type b conjugate vaccine polysaccharide derivative and conjugate bulk,group A and C meningococcal conjugate vaccine polysaccharide derivative and conjugate bulk were 97.4%,102.4%,96.8% and 99.8% respectively,with all CV values less than 15%;In the range of 0.312 5 ~ 80 ng/mL,cyanogroup concentration showed a good linear relationship with peak height with a regression equation of y = 133.13 x + 57.556,R^(2)= 0.999 1;The LOD was 0.2 ng/mL,and the LOQ was 0.6 ng/mL.The average recoveries of the control solution were 108.9%,106.5%,103.5% respectively with RSD value of 6.4% in the groups added with 5,10 and 20 ng/mL of cyanogroup and the CV values of precision verification were all less than 15%;The average concentrations of the control solution injected 20 times continuously were 76,38,18and 5 ng/mL with all CV values less than 15%,when cyanogroup concentration was 80,40,20 and 5 ng/mL,respectively.Cyanide residues were detected in 13 batches of derivative bulk samp

关 键 词：连续流动注射安培法 多糖蛋白结合疫苗 氰化物 A群C群脑膜炎球菌多糖结合疫苗 B型流感嗜血杆菌结合疫苗 

分 类 号：R392-3[医药卫生—免疫学]