高效液相色谱-电雾式检测器测定疫苗原液中去氧胆酸钠残留量  

Determination of Residual Sodium Deoxycholate in Vaccine Bulk by HPLC-CAD

作　　者：邵天舒 李珉 李玉立 李潇 李文东 SHAO Tianshu;LI Min;LI Yuli;LI Xiao;LI Wendong(Beijing Institute for Drug Control,NMPA Key Laboratory for Safety Research and Evaluation of Innovative Drug,Beijing 102206,China)

机构地区：[1]北京市药品检验研究院,国家药品监督管理局创新药物安全研究与评价重点实验室,北京102206

出　　处：《医药导报》2023年第3期401-404,共4页Herald of Medicine

基　　金：北京市科技新星计划(Z211100002121007)。

摘　　要：目的建立检测甲型肝炎灭活疫苗和23价肺炎球菌多糖疫苗原液中去氧胆酸钠残留量的高效液相色谱-电雾式检测器方法。方法采用安捷伦SB-C 18色谱柱(4.6 mm×250 mm,5μm),流动相为1%甲酸溶液-1%甲酸乙腈溶液,梯度洗脱,柱温30℃,流速1.0 mL·min^(-1),进样量50μL,电雾式检测器雾化器温度60℃,数据采集频率10 Hz,过滤常数3.6 s。结果样品中其他组分与去氧胆酸钠分离良好,对测定无干扰。去氧胆酸钠在0.475~47.50μg·mL^(-1)浓度范围内浓度与峰面积呈良好的线性关系(r=0.9990);甲型肝炎灭活疫苗及23价肺炎球菌多糖疫苗原液去氧胆酸钠含量平均回收率分别为100.1%和100.6%;去氧胆酸钠定量限约0.24μg·mL^(-1),检出限约0.12μg·mL^(-1)。结论该方法专属性好,灵敏度高,简便快捷,适用于疫苗原液中去氧胆酸钠残留量的检测。Objective To develop and verify a method for determining residual sodium deoxycholate in the bulk of inactivated hepatitis A vaccine and 23-valent pneumococcal vaccine by high performance liquid chromatography(HPLC)coupled with charge aerosol detection(CAD).Methods An Agilent SB-C 18 column(4.6 mm×250 mm,5μm)with a column temperature of 30℃was adopted.The mobile phase was composed of water containing 1%formic acid and acetonitrile containing 1%formic acid,while the gradient eluted at a flow rate of 1.0 mL·min^(-1).The injection volume was 50μL.The evaporating temperature of CAD was 60℃,while the collection rate was 10 Hz,and the filter constant was 3.6 s.Results Any other peaks were separated entirely from the peak of sodium deoxycholate and showed no interference with the determination result.The linear range of sodium deoxycholate was 0.475-47.50μg·mL^(-1),with correlation coefficients(r)of 0.9990.The average results of spike recovery tests of sodium deoxycholate content in two vaccines were 100.1%and 100.6%,respectively.The LOQ and LOD were about 0.24μg·mL^(-1)and 0.12μg·mL^(-1),respectively.Conclusion The developed method has reasonable specificity and high sensitivity.Moreover,it was convenient and rapid,suitable for determining residual sodium deoxycholate in the bulk of the vaccine.

关 键 词：疫苗原液 去氧胆酸钠残留量 高效液相色谱 电雾式检测器 

分 类 号：R917[医药卫生—药物分析学] R186[医药卫生—药学]