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  

Determination of ribose content in Haemophilus influenza type b polysaccharide by nuclear magnetic resonance

作　　者：普元倩 朱文勇[2] 寸怡娜[2] 代小虎[2] 张新文[2] 钟汉斌[2] 宋绍辉[2] 吴亚楠[2] 赵娜娜[1] 王明清[1,2] 廖国阳[2] 李卫东[2] 

机构地区：[1]昆明医科大学,云南昆明650550 [2]中国医学科学院北京协和医学院医学生物学研究所,云南昆明650118

出　　处：《中国生物制品学杂志》2014年第8期1081-1085,共5页Chinese Journal of Biologicals

基　　金：国家国际科技合作项目(2010DFB32890);Bill & Melinda Gates foundation(OPP1049425)

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。Objective To develop a method for determination of ribose content in Haemophilus influenzae type b polysaccharide by nuclear magnetic resonance（NMR）. Methods D-ribose at various concentrations were analyzed by the quantitative function of1H-NMR. The developed method was verified for precision and accuracy,and compared with the traditional chemical analysis. Results The relative standard deviations（RSD）of results of quantitative NMR were less than 1. 0%,while the relative errors,except that of samples at a concentration of 20 mmol / L（7. 4%）,were 1% - 3%.The recovery rates of samples at various concentrations were 97. 2% - 107%. However,the relative standard deviations（RSDs）of results of chemical analysis were 0. 6% - 7. 9%,while the relative errors were 3% - 6. 5%,and the recovery rates of samples were 93. 5% - 105. 3%. Conclusion The samples for quantitative NMR method are in a small quantity and may be quantified directly. With high precision,reproducibility and accuracy,the method may be used for quantitation of ribose in quality control of bacterial polysaccharide,by which no polysaccharide structure is destroyed.

关 键 词：核磁共振波谱法 b型流感嗜血杆菌多糖 含量测定 

分 类 号：R378.41[医药卫生—病原生物学] O657.61[医药卫生—基础医学]