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Quantitation of low level unconjugated polysaccharide in tetanus toxoid-conjugate vaccine by HPAEC/PAD following rapid separation by deoxycholate/HCl

Authors
Journal
Journal of Pharmaceutical and Biomedical Analysis
0731-7085
Publisher
Elsevier
Publication Date
Volume
21
Issue
6
Identifiers
DOI: 10.1016/s0731-7085(99)00183-1
Keywords
  • Anion Exchange Chromatography
  • Polyribosyl Ribitol Phosphate
  • Polysaccharide Conjugate Vaccine
  • Haemophilus Influenzae
  • Deoxycholate Precipitation

Abstract

Abstract A simple and rapid acid precipitation method has been applied successfully for separating free capsular polysaccharide of Haemophilus influenzae type b (polyribosyl ribitol phosphate, PRP) from PRP–tetanus toxoid conjugate (PRP-T) in a final dosage amount of low-level materials. The unconjugated PRP was found to stay in the supernatant without precipitation, while conjugated PRP-T was fully precipitated. High performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) has been applied for analysis of the PRP content in the supernatant after the separation. This method requires minimum sample handling and is specific, sensitive and reproducible making it suitable for release and stability testing of PRP-T in final containers.

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