离子色谱法测定脑膜炎球菌多糖蛋白结合疫苗中的高碘酸钠含量  

Determination of sodium periodate content in meningococcal polysac-charide protein conjugate vaccine by anion exchange chromatography

作　　者：傅元欣[1] 马庆华[1] 冯潇[1] 雒丽红[1] 魏然[1] 高雪军[1] 

机构地区：[1]兰州生物制品研究所有限责任公司,甘肃兰州730046

出　　处：《中国生物制品学杂志》2014年第1期103-108,共6页Chinese Journal of Biologicals

摘　　要：目的建立脑膜炎球菌多糖蛋白结合疫苗中残余高碘酸钠含量的检测方法。方法通过抗坏血酸将高碘酸根还原成碘离子,采用IonPac AS11-HC阴离子交换分析柱(4 mm×250 mm),上样量25μl,以50 mmol/LNaOH淋洗液等度洗脱,流速1.5 ml/min,电化学检测器测定还原出的I-,用Chromeleon色谱工作站记录并分析数据。对建立的方法进行专属性、准确性、重复性验证,确定该方法的最低检出限和最小定量限,并进行初步应用。结果等浓度的抗坏血酸和高碘酸钠的反应比例为4∶1时,高碘酸根可完全还原为I-。对照品I-在10μg/L^1 mg/L范围内,I-浓度和色谱峰面积呈良好的线性关系,r均大于0.999,回收率为91.66%~110.20%,RSD为0.2%~2.3%,最小检出限为1μg/L(信噪比3∶1),最小定量限为5μg/L(信噪比10∶1)。用建立的方法检测A群、C群、Y群和W135群脑膜炎球菌多糖蛋白结合物各2批,均未检测出高碘酸钠。结论离子色谱法可检测脑膜炎球菌多糖蛋白结合疫苗中的高碘酸钠含量,该方法操作简便、灵敏、快速,干扰小,重现性好,适用于对疫苗生产过程中的质量控制。Objective To develop a method for determination of residual sodium periodate （NalO4） content in meningo- coccal polysaecharide protein conjugate vaccine. Methods Periodate was reduced to iodide ion with aseorbic acid, then loaded onto IonPaeASll-HC anion exchange column （4 mm× 250 mm） at a loading of 25 μl, eluted with 50 mmol/L sodium hydroxide at a flow rate of 1. 5 ml / min, and determined with electrochemical detector, of which the data were recorded and analyzed by using Chromeleon chromatography workstation. The developed method was verified for specifici- ty, accuracy and reproducibility, determined for minimum detection and quantitative detection limits, and applied pre- liminarily. Results Periodate was completely reduced to iodide ion when the proportion of aseorbic acid to sodium perio- date, at equal concentrations, was 4 ： 1. The iodide ion concentration of control at a range of 10 μg / L - 1 mg / L showed good liner relationship to chromatographic peak area, with a r value of more than 0. 999. The recovery rate, RSD, minimum detection limit and minimum quantitative detection limit of the developed method were 91. 66% 110. 20%, 0. 2% - 2. 3%, 1 p.g/L [with a signal-to-noise ratio （SNR） of 3 ： 1] and 5 μg/L （with a SNR of 10 ： 1） respectively. Each of two batches of meningococeal polysaecharide protein conjugate vaccine of groups A, C, Y and W135 were determined by the developed method, and no sodium periodate was detected. Conclusion The developed method was simple, sensitive, rapid, with little interference and good reproducibility, which was suitable for the quality control of vaccine during production.

关 键 词：离子色谱法 脑膜炎球菌多糖蛋白结合疫苗 高碘酸钠 含量测定 

分 类 号：O657.75[理学—分析化学] R392.33[理学—化学]