Abstract

RICO, Jannete; MERCHAN, Yaima; CUEVAS, Matilde  and  MARQUEZ, Jenny. Validation of a method of polysaccharide traces determination by HPLC-FL in Outer Membrane Vesicles from N. meningitidis. Vaccimonitor [online]. 2010, vol.19, n.2, pp. 18-23. ISSN 1025-0298.

The High-Resolution Liquid Chromatography-Fluorescence has proven to be a sensitive tool for trace detection in pharmaceuticals and it is frequently used for the quantification of some bacterial polysaccharides that have sialic acid as repeating unit. In this paper we aim at establishing and validating HPLC-fluorescence to determine polysaccharide content of residual B outer membrane vesicles from Neisseria meningitidis. We assembled the HPLC-FL method to the quantification of this residue, using a C18 reverse phase column (ODS UltraSphere, Beckman, USA). We evaluated six lots of outer membrane vesicles from different strains Cu 385/83 and NZ 228. It was demonstrated that the HPLC-FL method allows the quantification of polysaccharide B in a specific and exact manner, and that it complied with the validation parameters for a method for determining trace amounts by HPLC.

Keywords : HPLC-FL; validation; polysaccharide B.

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