疫苗中游离甲醛含量新型高通量检测方法的建立  

Development of a novel high-throughput method for the determination of free formaldehyde content in vaccines

作　　者：李玉立 李珉 马一星 纪宏 朱晓月 何欢 李文东 LI Yu-li;LI Min;MA Yi-xing;JI Hong;ZHU Xiao-yue;HE huan;LI Wen-dong(Beijing Institute for Drug Control(Beijing Center for Vaccine Control),NMPA Key Laboratory for Research and Evaluation of Generic Drug,Beijing Key Laboratory of Analysis and Evaluation on Chinese Medicine,Beijing 102206,China)

机构地区：[1]北京市药品检验研究院(北京市疫苗检验中心),国家药品监督管理局仿制药研究与评价重点实验室,中药成分分析与生物评价北京市重点实验室,北京102206

出　　处：《药物分析杂志》2024年第8期1365-1372,共8页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立一种新型高通量检测方法,用于测定疫苗中游离甲醛含量,并与2020年版《中华人民共和国药典》(简称《中国药典》)方法进行结果比对。方法:疫苗样品不经任何前处理,直接加入顶空瓶中与衍生化试剂1%对甲基苯磺酸-乙醇溶液(1%TsOH-EtOH溶液)混合后进行测定。考察了衍生化试剂浓度、用量及最佳衍生化条件(顶空孵育温度和时间)。采用DB-624气相毛细管柱,顶空进样,程序升温,检测器为氢离子火焰检测器(FID)。结果:线性范围为0.25~100μg·mL^(-1)(r=0.9995),检测限和定量限分别为0.10μg·mL^(-1)和0.25μg·mL^(-1);多种疫苗加样回收率均在94%以上,RSD均低于6%,专属性、精密度及耐用性均符合要求。多批次检测结果均与药典方法测定结果无显著差异。结论:开发并验证了1种新型衍生化试剂(1%TsOH-EtOH溶液)用于疫苗中游离甲醛含量的测定,简捷高效,并可实现高通量检测。该方法既可以解决2020年版《中国药典》方法中的多种溶液配制,检验步骤多以及检验周期较长的问题,又可以解决独立衍生化前处理操作带来的工作量的增加与检验效率的降低,可用于疫苗等生物制品复杂基质中的游离甲醛的测定,对于化学药品、中药及辅料中的游离甲醛的测定也有参考意义。Objective:To develop a novel and efficient assay for the determination of free formaldehyde content in vaccines,and to compare the results with the method of the Chinese Pharmacopoeia(ChP)2020 edition.Methods:Vaccine samples were added directly into headspace vials without any pretreatment and tested after mixed with a new derivatization reagent,1%p-toluenesulfonic acid-ethanol solution(1%TsOH-EtOH solution).The concentration of the reagent,the dosage and the optimal derivatization conditions(headspace incubation temperature and time)were tested and confirmed.A DB-624 gas capillary column was used with temperature programming.The detector was a hydrogen-ion flame detector(FID)and the sample was injected in headspace.Results:The linear range was 0.25-100μg·mL^(-1)(r=0.9995),the detection limit and the quantification limit were 0.10μg·mL^(-1) and 0.25μg·mL^(-1),respectively.The recovery rates of several kinds of vaccines were all over 94%,and the RSDs were all less than 6%,which were in line with the requirements of specificity,precision and durability.There were no significant differences between the results of multiple batches and those determined by ChP 2020 methods.Conclusion:In this paper,a novel derivatization reagent(1%TsOH-EtOH solution)was developed and validated for the determination of free formaldehyde content in vaccines,which is simple,efficient and can achieve high-throughput detection.The method can just solve the problems in other current detection methods.The method of derivatization headspace gas chromatography for the determination of free formaldehyde content is applicable to vaccines and other biologics,and provide a reference for chemicals,traditional Chinese medicine and excipients.

关 键 词：疫苗 顶空气相色谱法 游离甲醛 衍生化 高通量 药典方法比对 

分 类 号：R917[医药卫生—药物分析学]