气相色谱法测定甲型H1N1流感病毒裂解疫苗中乙醚的残留量  

Determination of residual ether content in influenza A H1N1 split vaccine by gas chromatography

作　　者：吴金妍[1] 程鹏飞[1] 秦洁[1] 郁佳俊[1] 

机构地区：[1]上海生物制品研究所有限责任公司,上海200052

出　　处：《中国生物制品学杂志》2014年第3期412-414,418,共4页Chinese Journal of Biologicals

摘　　要：目的目的采用气相色谱法测定甲型H1N1流感病毒裂解疫苗中乙醚的残留量，并对该方法进行验证及初步应用。方法采用气相色谱法定量测定甲型H1N1流感病毒裂解疫苗中的乙醚含量，顶空条件：平衡温度为90℃，平衡时间为30min。色谱条件：色谱柱：HP-FFAP毛细管色谱柱（25m×0．32mm×0．5μm）；载气：氮气；流量：1．0-2．0ml／min；进样口温度：160℃；检测器温度：250℃；柱温：45℃；分流比：5：1；进样体积：1ml。对建立的方法进行验证及初步应用，并建立质量标准。结果该方法检测乙醚残留量溶剂对所测组分无干扰；系统适应性良好；乙醚浓度在11-110μg／ml范围内，标准曲线的线性关系良好（r=0．9993）；最低定量限为0．028μg／ml；检测对照品溶液的相对标准偏差（尼如）为2．98％；加标回收率在83．08％-87．58％之间。应用该方法检测9批疫苗样品中的乙醚残留量在0．00003％-0．00055％之间，符合《中国药典》二部（2005版）附录VIP应不高于0．5％的要求。将乙醚残留量作为该疫苗半成品的质量标准检测项目，限度定为0．05％。结论气相色谱法测定甲型H1N1流感病毒裂解疫苗中乙醚的残留量系统适用性、线性、精密性和准确性良好，且易于操作，简便快速，可用于甲型H1N1流感病毒裂解疫苗中乙醚残留量的测定。Objective To determine the residual ether content in influenza A H1N1 split vaccine by gas chromatography （GC）, and verify and preliminarily apply the developed method. Methods The residual ether content in influenza A H 1N 1 split vaccine was developed by GC. The condition for equilibration was as follows： temperature： 90 °C; time： 30 min. The condition for GC was as follows： chromatographic column： HP-FFAP capillary chromatographic column （25 m x 0. 32 mmx0. 5 μm）; carrier gas： nitrogen; flow rate： 1. 0 - 2. 0 ml/min; temperature of sample entry： 160 °C; temperature of detector： 250 °C; temperature of column： 45 ~C; split ratio： 5 ： 1; sample loading： 1 ml. The developed method was verified and preliminarily applied, based on which a quality standard was established. Results By the developed method, the solvent for determination of residual ether content showed no interference to the component to be tested. The method showed good systemic suitability. The standard curve showed good linearity at an ether concentration of 11 - 110 μg/ml （r = 0. 999 3 ）. The minimum quantification limit of the developed method was 0. 028 μg/ml. The relative standard deviation （RSD） of determination result of control solution was 2. 98%. The spike recovery rate was 83. 08% - 87. 58%. The residual ether contents in nine batches of influenza A H1N1 split vaccine determined by the developed method were 0. 000 03% - O. 000 55%, which met the requirements in Appendix Ⅷ P of Chinese Pharmacopoeia （Volume Ⅱ , 2005 edition） （not more than 0. 5% ）. Conclusion The developed GC method showed high systemic suitability, linearity, precision and accuracy, which was easy to handle, simple, rapid, and was suitable for determination of residual ether in influenza A H1N1 split vaccine

关 键 词：流感病毒A型 H1N1亚型 疫苗 乙醚 气相色谱法 

分 类 号：R373.13[医药卫生—病原生物学] R392-33[医药卫生—基础医学]