Abstract：
A selective and sensitive method was evaluated for quantitation of meningococcal X (Men X) polysaccharide in pentavalent meningococcal A, C, W, Y and X conjugate vaccine using different acid hydrolysis conditions like HCl, TFA, HF, HF-TFA, and HF-HCl. High-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) using CarboPac PA10 column was used to identify the hydrolyzed products based on retention time and its comparison with monosaccharide standards. Complete release of glucosamine (GlcN) from Men X in monovalent bulk and pentavalent vaccine samples was achieved using HF hydrolysis at 80 °C for 2 h. The Men X HF-hydrolyzed polysaccharide to glucosamine along with the reference standard was identified using collision-induced dissociation (CID) electrospray mass spectroscopy and the MS/MS fragments of m/z 162, m/z 144 and m/z 84. Meningococcal polysaccharide concentration was determined with a correlation coefficient r2 >0.99 using polysaccharide reference standard. The serogroups A, W, and Y were converted to their monosaccharides units and quantified using this method however, milder acid hydrolysis 0.1 M HCl 80 °C 2 h for release of sialic acid for Men C polysaccharide was found to be more suitable. These methods will provide necessary tools and prove to be beneficial to laboratories developing new saccharide-based vaccine combinations.
摘要：
评估了一种选择性和灵敏的方法，用于定量五价脑膜炎球菌A中的脑膜炎球菌X（MenX）多糖，C,W,Y和X结合疫苗使用不同的酸水解条件，如HCl，TFA,HF,HF-TFA,和HF-HCl。使用CarboPacPA10柱的具有脉冲安培检测的高效阴离子交换色谱（HPAEC-PAD）用于根据保留时间及其与单糖标准品的比较来鉴定水解产物。使用HF在80°C下水解2小时，从单价批量和五价疫苗样品中完全释放葡糖胺（GlcN）。使用碰撞诱导解离（CID）电喷雾质谱和m/z162，m/z144和m/z84的MS/MS片段鉴定了MenXHF水解的多糖与参考标准。使用多糖参考标准以相关系数r2>0.99测定脑膜炎球菌多糖浓度。血清群A,W,和Y被转化为它们的单糖单元，并使用这种方法进行定量，较温和的酸水解0.1MHCl80°C2h释放唾液酸对于MenC多糖更合适。这些方法将提供必要的工具，并证明有利于实验室开发新的基于糖的疫苗组合。
