离子色谱法检测Y群和W135群脑膜炎球菌荚膜多糖含量

Content determination of groups Y and W135 meningococcal capsule polysaccharides by high-performance anion-exchange chromatography with pulsed amperometric detection

目的：建立Y群W135群脑膜炎球菌荚膜多糖含量的高效阴离子交换色谱-脉冲安培检测法（high-performance anion-exchange chromatography with pulsed amperometric detection，HPAEC—PAD），并对该方法进行验证。方法：通过对水解温度、水解时间和水解用酸及其浓度的筛选，确定最佳的水解条件；利用固总法和化学法衡量HPAEC．PAD法的准确性，检测ACYW135群脑膜炎球菌多糖疫苗和Y群W135群多糖蛋白结合疫苗原液，评价其重复性和精密性。结果：最佳水解条件为2mol·L^-1 HCL 80℃水解1h。色谱法的检测结果与固总法和化学法比较，CV在1．60％～5．04％，结果准确。检测Y群和W135群多糖含量重复性和精密性均良好。结论：HPAEC-PAD法可以用于Y群W135群多糖含量的测定。

Objective： To determine and verify contents of groups Y and W135 meningococcal capsule pol- ysaccharides by high-performance anion-exchange chromatography with pulsed amperometric detection （HPAEC- PAD）. Methods： The best hydrolyzed condition was confirmed by optimizing hydrolyzed temperature, time and concentration. The accuracy was evaluated by a total solid method and a colorimetry method. The reproducibility and accuracy were assessed by determination contents of ACYW135 polysaccharide vaccine and polysaccharide conjugate vaccine. Results： The best hydrolyzed condition was 2 mol·L^-1 hydrochloric acid at 80 ℃ for 1 h. Compared with the total solid method and colorimetry method, the CV was 1.60% - 5.04% by HPAEC-PAD method. It had high reproducibility and accuracy in determining contents of group Y and group W135 polysaccharides. Conclusion： The HPAEC-PAD method can be used for content determination of group Y and group W135 polysaccharides.