title: HPAEC-PAD method for the analysis of alkaline hydrolyzates of Haemophilus influenzae type b capsular polysaccharide.
abstract: A gradient method has been devised for the rapid analysis of alkaline hydrolyzates of Haemophilus influenzae type b (Hib) capsular polysaccharide-based vaccines by high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). As compared with published procedures, peak shape and sensitivity were significantly improved with this approach, analysis time was short and there was little interference from impurities. The limits of detection and quantification were established with a purified reference polysaccharide. We propose this method as a practical alternative for the analysis of minute amounts of Hib polysaccharide, which can be lower than with the conventional approaches.
authors.affiliation: Unit Vaccinology, Centre for Infectious Disease Control (CIb), National Institute for Public Health and the Environment (RIVM), P.O. Box 1, 3720 BA, Bilthoven, The Netherlands.
authors.forename: Alex
authors.initials: A
authors.lastname: de Haan
authors.forename: Robert M F
authors.initials: RM
authors.lastname: van der Put
authors.forename: Michel
authors.initials: M
authors.lastname: Beurret
mesh_terms: D016667:Bacterial Capsules / Q000737:chemistry* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D018073:Haemophilus Vaccines / Q000737:chemistry; D019741:Haemophilus influenzae type b / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D012680:Sensitivity and Specificity; D019346:Sodium Acetate / Q000737:chemistry; D012972:Sodium Hydroxide / Q000737:chemistry
chemical_list: D018073:Haemophilus Vaccines; D019346:Sodium Acetate; D012972:Sodium Hydroxide
keywords: Haemophilus influenzae type b (Hib); alkaline hydrolysis; conjugate vaccine analysis; high performance anion exchange chromatography (HPAEC); polysaccharide analysis; pulsed amperometric detection (PAD)