Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China; Jiangnan University-Beijing Minhai Biotechnology Joint Research Center of Carbohydrate-Based Vaccines, Jiangnan University, Wuxi 214122, China.
National & Local Joint Engineering Laboratory for Novel Vaccine Development Technology, Beijing Minhai Biotechnology Co., Ltd., Beijing 102600, China; Jiangnan University-Beijing Minhai Biotechnology Joint Research Center of Carbohydrate-Based Vaccines, Jiangnan University, Wuxi 214122, China.
Chin J Nat Med. 2022 Aug;20(8):633-640. doi: 10.1016/S1875-5364(22)60184-5.
The ribose and phosphorus contents in Haemophilus influenzae type b (Hib) capsular polysaccharide (CPS) are two important chemical indexes for the development and quality control of Hib conjugate vaccine. A quantitative H- and P-NMR method using a single internal standard was developed for simultaneous determination of ribose and phosphorus contents in Hib CPS. Hexamethylphosphoramide (HMPA) was successfully utilized as an internal standard in quantitative H-NMR method for ribose content determination. The ribose and phosphorus contents were found to be affected by the concentration of polysaccharide solution. Thus, 15-20 mg·L was the optimal concentration range of Hib CPS in DO solution for determination of ribose and phosphorus contents by this method. The ribose and phosphorus contents obtained by the quantitative NMR were consistent with those obtained by traditional chemical methods. In conclusion, this quantitative H- and P-NMR method using a single internal standard shows good specificity, accuracy and precision, providing a valuable approach for the quality control of Hib glycoconjugate vaccines.
b 型流感嗜血杆菌（Hib）荚膜多糖（CPS）的核糖和磷含量是 Hib 结合疫苗开发和质量控制的两个重要化学指标。建立了一种使用单一内标物的定量 H 和 P-NMR 方法，用于同时测定 Hib CPS 中的核糖和磷含量。六甲基磷酰胺（HMPA）成功地用作定量 H-NMR 法中测定核糖含量的内标物。发现核糖和磷含量受多糖溶液浓度的影响。因此，对于通过该方法测定核糖和磷含量，Hib CPS 在 DO 溶液中的最佳浓度范围为 15-20mg·L。通过定量 NMR 获得的核糖和磷含量与传统化学方法获得的核糖和磷含量一致。总之，这种使用单一内标物的定量 H 和 P-NMR 方法具有良好的特异性、准确性和精密度，为 Hib 糖缀合物疫苗的质量控制提供了有价值的方法。
