摘要The ribose and phosphorus contents in Haemophilus influenzae type b(Hib)capsular polysaccharide(CPS)are two im-portant chemical indexes for the development and quality control of Hib conjugate vaccine.A quantitative1H-and 31P-NMR method using a single internal standard was developed for simultaneous determination of ribose and phosphorus contents in Hib CPS.Hexa-methylphosphoramide(HMPA)was successfully utilized as an internal standard in quantitative 1H-NMR method for ribose content de-termination.The ribose and phosphorus contents were found to be affected by the concentration of polysaccharide solution.Thus,15-20 mg·mL-1 was the optimal concentration range of Hib CPS in D2O 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 tradi-tional chemical methods.In conclusion,this quantitative1H-and 31P-NMR method using a single internal standard shows good spe-cificity,accuracy and precision,providing a valuable approach for the quality control of Hib glycoconjugate vaccines.