摘要
目的用高效阴离子交换色谱-脉冲安培检测法(HPAEC-PAD)测定A、C、Y和W135群脑膜炎球菌多糖含量,并对该方法进行验证及初步应用。方法采用Carbo Pac~?PA-1 4×250 mm分析柱与Aminotrap 4×50 mm保护柱;使用氢氧化钠/醋酸钠梯度洗脱,以多糖抗原标准溶液质量浓度为横坐标,其相应的峰面积为纵坐标,绘制标准曲线,并对建立的方法进行专属性、精密度和准确性验证;同时与传统方法的检测结果进行比较分析。结果 A、C、Y和W135群脑膜炎球菌多糖质量浓度在0.5～27.0μg/m L范围内与峰面积均具有良好线性关系,r=0.999。该方法专属性高,分析结果重复性良好,RSD均<5%;回收率均在95%～105%范围内;与传统的火箭免疫电泳法相比,检测结果差异均无统计学意义(P>0.05)。结论 HPAEC-PAD准确、可靠,重复性好,可用于检测脑膜炎球菌多糖原液及疫苗成品中的多糖含量。
Objective The group A,C,Y and W135 meningococcal polysaccharide contents were detected by a high performance anion exchange chromatography with pulse amperometrydetector（ HPAEC-PAD）,which was verified and priminary applied. Methods Carbo Pac C PA-1 4 x 250 mm and Aminotrap 4 x 50 mm（ 046122） were used as the analytical and guard column,then the gradient elution was conducted by using solution of sodium hydroxide/sodium acetate.The polysaccharide content and its corresponding peak area were used as the abscissa and ordinate,and the standard curves were set up. The developed method was verified for specificity,accuracy and precision. Furthermore,the results were compared with those of the tradition assay. Results The detection showed that a good linearity was found in the range of 0.5～ 27.0 μg/m L polysaccharides and the corresponding peak area,with r = 0.999. The developed HPAEC-PAD were found in advantagies with high specificity,and good repeatability（ RSD〈5%）. The recovery rate of polysaccharide was in the range of 95% ～ 105%. Compared with traditional rocket immuno-electrophoresis method,the results from HPAEC-PAD had no a statistical difference（ P〉0.05）. Conclusion This method is accurate and reliable,which could be used in detection of meningococcal polysaccharide bulk and the polysaccharide content in relevant vaccine.
作者
李亚南
赵丹
徐颖华
毛琦琦
陈苏京
李茂光
叶强
LI Ya-nan;ZHAO Dan;XU Ying-hua;MAO Qi-qi;CHENG Su-jing;LI Mao-guang;YE Qiang(Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products,National Institute for Food and Drug Control,Beifing 102629,China)
出处
《微生物学免疫学进展》
2018年第5期44-48,共5页
Progress In Microbiology and Immunology
关键词
阴离子交换色谱法
脉冲安培检测器
脑膜炎球菌多糖
Anion exchange chromatography
Pulse amperometrydetector
Meningococcal polysaccharides
