Development and Verification of Phosphorus Assay in Haemophilus Inflluen-zae Type B Conjugate Vaccine
Objective:To establish and verify an ultrafiltration method combined with ultraviolet spec-trophotometry for detection of phosphorus contents in Haemophilus inflluenzae type b(Hib)conjugate vac-cines.Methods:The sample was dissolved with deionized water and then ultrafiltrated.Hib polysaccharide in the ultrafiltration tube was collected.The sample was digested with perchloric acid and sulfuric acid(1:1).Ammonium molybdate was used to form phosphomolybdic acid with phosphorus under acidic conditions,and then blue compound was formed with vitamin C with maximum absorption at 825 nm.The linearity,precision,stability,and accuracy of this method were verified.The established method was used to detect the phosphorus contents of three commercial batches of freeze-dried Hib conjugate vaccines.Results:The phosphorus concentrations at a range of 1-10 μg·mL-1 showed good linear relationship,the coefficient r was 1.000,the linear regression equation was Y=8.878 × 10-2 X+2.402 × 10-3.The average recovery at low,medium,and high concentration levels were 97.35％,96.65％and 97.42％(n=3),respectively,with RSD of 2.55％,0.95％and 0.95％.The RSD of the reproducibility test was 2.81％,and the intermediate precision RSD was 2.14％.The derivative solutions of phosphorus standard were stable at room temperature for 1 hour with RSD at 0.39％.Conclusion:This method has accurate and reliable results,and can be used for the detection of phosphorus contents in freeze-dried Haemophilus influenzae type b conjugate vaccines.It also provides clues for the determination of phosphorus contents in other polysaccharide protein conjugate vaccines.
Ultrafiltration methodFreeze-dried Haemophilus inflluenzae type b conjugate vaccinePhosphorusAscorbic acid
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维普
