离子色谱法测定脑膜炎球菌多糖蛋白结合疫苗中的1,4-丁二胺含量

Determination of 1,4-butanediamine content in meningococcal polysaccharide protein conjugate vaccine by ion chromatography

目的采用离子色谱法测定脑膜炎球菌多糖蛋白结合疫苗中的1,4-丁二胺含量,并对方法进行验证。方法采用Ion Pac CS17(4 mm×250 mm)分析柱,上样量25μl,以10 mmol/L甲基磺酸淋洗液进行等度洗脱,流速1.0 ml/min,电导检测器检测,Chromeleon色谱工作站记录并分析数据。对该方法进行专属性、线性、准确性、重复性验证,并确定该方法的最低检出限和最小定量限。结果该方法检测1,4-丁二胺专属性较强;在40μg/L^1 mg/L范围内,1,4-丁二胺浓度与色谱峰面积呈良好的线性关系,r2均大于0.999;向A、C、W135、Y群脑膜炎球菌多糖蛋白结合物原液中分别添加低、中、高浓度的1,4-丁二胺,重复测定5次的回收率在97.23%~104.94%之间,重复测定10次的相对标准偏差(RSD)在0.8%~1.8%之间;该方法的最低检出限为5μg/L(信噪比3∶1),最小定量限为40μg/L(信噪比10∶1)。结论离子色谱法简便、快速,灵敏度高,可用于检测脑膜炎球菌多糖蛋白结合疫苗中1,4-丁二胺的含量。

Objective To determine the 1,4-butanediamine content in meningococcal polysaccharide protein conjugate vaccine by ion chromatography and verify the method. Methods Ion Pac CS17 analytical column（4 mm × 250 mm） was used at a loading of 25 μl, eluted with 10 mmol / L methanesulfonic acid at a flow rate of 1. 0 ml / min, and determined with electrolytic conductivity detector, of which the data were recorded and analyzed by using Chromeleon chromatography workstation. The developed method was verified for specificity, linearity, accuracy and reproducibility, and determined for minimum detection and quantitative detection limits. Results The developed method showed high specificity for determination of 1,4-butanediamine content, and showed good linear relationship to chromatographic peak area at a concentration range of 40 μg / L ~ 1 mg / L, with r2 values of more than 0. 999. The bulk of groups A, C, W135 and Y meningococcal polysaccharide protein conjugate vaccine was added with 1,4-butanediamine at low, moderate and high concentrations, of which the recovery rates in five tests were 97. 23% ~ 104. 94%, and the RSDs of determination results in ten tests were 0. 8% ~ 1. 8%. The minimum detection limit and minimum quantitative detection limit of the developed method were 5[with a signal-to-noise ratio（SNR）of 3 ∶ 1]and 20 μg / L（with a SNR of 10 ∶ 1）respectively. Conclusion Ion chromatography was simple, rapid and sensitive, which might be used for determination of 1,4-butanediamine content in Meningococcal polysaccharide protein conjugate vaccine.