顶空气相色谱法测定四价脑膜炎球菌多糖蛋白结合疫苗中残余二氧六环

Determination of the residual content of dioxane in tetravalent meningococcal polysaccharide protein conjugate vaccine by head-space gas chromatography

目的建立并验证四价脑膜炎球菌多糖蛋白结合疫苗中残余二氧六环的检测方法。方法采用顶空气相色谱法,样品中加入碳酸钠固体,产生"盐析"效应,以增加二氧六环在顶空瓶中的浓度。通过HP-5毛细管柱(30 m×0.32 mm×0.25μm,5%Phenyl Methyl-Siloxane),氢火焰检测器测定四价脑膜炎球菌多糖蛋白结合疫苗中残余二氧六环。结果二氧六环平均回收率及变异系数分别为86.16%～106.54%和2.4%～8.7%,在0.5004～40.032mg/L范围内相关系数值r=0.999 41,线性关系好,最低检测限为0.5 mg/L,最低定量限为2.5 mg/L。结论该方法溶剂用量少、灵敏度高、出峰特异、专属性强、色谱峰型好、分离度高、出峰时间短、影响因素少,是一种易于实现仪器自动化的分析方法。

Objective To establish a method in determination of the residual dioxane in tetravalent meningococcal polysaccharide-protein conjugate vaccine.Methods Head-space gas chromatography was used in this test.When sodium carbonate was added in sample,the effect of salting-out resulted in increasing the concentration of dioxane in head-space bottles.Then the content of dioxane was analyzed by passing through a HP-5 capillary column（30 m×0.32 mm×0.25 μm,5% Phenyl Methyl-Siloxane） and in combination with a hydrogen flame detector.Results The average recovery of dioxane was 86.16%-106.54% and the average CV value was 2.4%-8.7%.A good liner correlation was obtained over the range of 0.5004-40.032 mg/L,r = 0.99941.The limit of detection was 0.5 mg/L,the method can quantify the dioxane content with a minimum amount of 2.5 mg/L for the meningococcal PS and has a sensitivity at least 10 times higher than that of the noise assay.Conclusions This method requires minimum solvent and is sensitive,specific and rapid,moreover,it can be readily adapted for use.