Evaluation of the Saccharide Content of the WHO 2nd International Standard for Haemophilus Influenzae Polysaccharide Polyribosyl Ribitol Phosphate (PRP) by HPAECPAD Analysis Following Acid Hydrolysis

• Published in: WHO drug information Vol. 36; no. 4; pp. 887 - 904
• Main Authors: Rosskopf, Ute, von Hunolstein, Christina, Gaggioli, Andrea
• Format: Journal Article
• Language: English
• Published: Geneva World Health Organization 01-10-2022
• Subjects: Acids; Anion exchange; Anion exchanging; Calibration; Carbohydrates; Chromatography; Collaboration; Electrical measurement; Hydrolysis; International standards; Laboratories; Moisture content; Polysaccharides; Reproducibility; Ribose; Test methods; Vaccines
• ISSN: 1010-9609; 1996-8361

With this report, we present the results of a collaborative study evaluating the content in mg per ampoule of the "WHO 2nd International Standard (IS) for Haemophilus influenzae polysaccharide Polyribosyl Ribitol Phosphate (PRP), NIBSC code: 12/306" (WHO 2nd IS for PRP), by applying High Performance Anion Exchange Chromatography with Pulsed Amperometric Detection (HPAEC-PAD) following acid hydrolysis of the PRP. Seventeen laboratories from eleven countries participated in the collaborative study. The outcome of this study revealed a PRP content of the WHO 2nd IS for PRP of 4.338 ± 0.203 mg/ampoule. Results of a previous collaborative study that included eleven participating laboratories, yielded a content of 4.904 ± 0.185 mg/ampoule when applying the ribose assays (i.e., by orcinol method). Based on the outcome of this study, using HPAEC-PAD and acid hydrolysis of PRP, it is concluded that the determination of the PRP content is method dependent and therefore assigned values for the WHO 2nd IS for PRP should be tailored according to the applied method.