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1 2000

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Identification, interaction and Kd studies of bacterial biopolymers by use of superose 6 gels and UV monitoring

Munoz E. C., Baldor C. T., Munoz M. E., Cabrera R. A., Infante J. F. B., Sierra G. G.

Acta Chromatographica

2000

No. 10

147-157

A spectrophotometric study has been performed on one of the bacterial polysaccharides most often used in vaccines – Neisseria meningi-tisdis group C polysaccharide – and the results were compared with those obtained for Haemophilus influenzae type b polysaccharide. The effect of mobile phase has been investigated and specific spectrophotometric absorption has been assessed to determine the feasibility of ultra-violet (UV) detection and quantification. The classical method established by the World Health Organisation (WHO) for analysis of these polysaccharides was compared with high-performance liquid chromatography (HPLC). The results obtained showed the possibility of using UV detection for polysaccharide quantification, and that there was good correlation between results obtained by use of the HPLC and WHO methods for characterization of molecular size; use of HPLC enabled more rapid and more accurate analysis. Biopolymers that usually occur with the polysaccharides were also studied by HPLC on Superose 6 gels with UV monitoring at 206 nm. Peaks of different compounds, including the polysaccharides, could be identified in one run.