核磁共振波谱法测定b型流感嗜血杆菌荚膜多糖中的核糖含量

发布时间：2018-04-03 23:46

本文选题：核磁共振波谱法　切入点：b型流感嗜血杆菌多糖　出处：《中国生物制品学杂志》2014年08期

【摘要】：目的建立核磁共振波谱(nuclear magnetic resonance,NMR)法测定b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)荚膜多糖中核糖含量的方法。方法利用NMR技术中的氢核磁共振波谱(1H-NMR)的定量功能,对不同浓度的D-核糖进行定量分析,验证该方法的精密度及准确度,并与传统化学方法进行比较。结果 NMR法定量结果相对标准偏差(RSD)均小于1.0%,相对误差除最低浓度20 mmol/L为7.4%外,其余浓度均在1%~3%之间,不同浓度的样品回收率在97.2%~107%之间;化学法定量结果 RSD在0.6%~7.9%之间,相对误差在3%~6.5%之间,不同浓度样品回收率在93.5%~105.3%之间;两种方法检测的3批Hib荚膜多糖中核糖含量无明显差别,均在《中国药典》三部(2010版)规定的320~410 mg/g之间。结论 1H-NMR定量方法取样少,可直接定量,该方法精密度高、准确度高、重现性好,且不破坏多糖结构,可应用于细菌荚膜多糖质量控制的核糖定量。
[Abstract]:Objective to establish a method for the determination of ribose in Haemophilus influenzae type bhib capsule polysaccharides by nuclear magnetic resonance spectroscopy (NMR).Methods using the quantitative function of 1H-NMRs in NMR, quantitative analysis of different concentrations of D-ribose was carried out. The precision and accuracy of the method were verified and compared with the traditional chemical method.Results the relative standard deviation (RSD) of NMR was less than 1.0. The relative error was between 1.0% and 7.4%, except the lowest concentration of 20 mmol/L was 7.4%. The recoveries of samples of different concentrations were between 97.2% and 10.7%, and the relative errors were between 0.6% and 7.9% by chemical method.The relative error was between 6.5% and 93.5%, and the recoveries of samples with different concentrations were between 93.5% and 105.3%. There was no significant difference in the contents of ribose in the three batches of Hib capsule polysaccharides detected by the two methods, both of which were in the range of 320,410 mg/g as stipulated in the Chinese Pharmacopoeia, three parts of Chinese Pharmacopoeia and three parts of Chinese Pharmacopoeia.Conclusion 1H-NMR quantitative method can be used in the quality control of bacterial capsule polysaccharides because of its high precision, high accuracy, good reproducibility and no damage to the structure of polysaccharides.
【作者单位】：昆明医科大学;中国医学科学院北京协和医学院医学生物学研究所;
【基金】：国家国际科技合作项目(2010DFB32890) Bill & Melinda Gates foundation(OPP1049425)
【分类号】：R378;O657.61

【相似文献】