Development of an on-line preconcentration method for the analysis of pathogenic lipopolysaccharides using capillary electrophoresis–electrospray mass spectrometry: Application to small colony isolates

The present investigation describes the use of on-line chromatographic preconcentration coupled to capillary zone electrophoresis–electrospray mass spectrometry (cPC–CZE–ES-MS) for trace level analysis of negatively charged lipopolysaccharides (LPS) obtained from pathogenic strains of Haemophilus in...

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Bibliographic Details
Published in:Journal of Chromatography A 1998-08, Vol.817 (1), p.325-336
Main Authors: Li, J, Thibault, P, Martin, A, Richards, J.C, Wakarchuk, W.W, van der Wilp, W
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Bacteriology
Biological and medical sciences
Carbohydrate Sequence
Electrophoresis, Capillary - methods
Electrospray mass spectrometry
Fundamental and applied biological sciences. Psychology
Haemophilus influenzae - chemistry
Lipopolysaccharides
Lipopolysaccharides - analysis
Lipopolysaccharides - chemistry
Mass Spectrometry - methods
Microbiology
Molecular Sequence Data
Morphology, structure, chemical composition
Peptides
Saccharides
Sensitivity and Specificity
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Summary:The present investigation describes the use of on-line chromatographic preconcentration coupled to capillary zone electrophoresis–electrospray mass spectrometry (cPC–CZE–ES-MS) for trace level analysis of negatively charged lipopolysaccharides (LPS) obtained from pathogenic strains of Haemophilus influenzae. The analytical performance of two different types of adsorption media [i.e., C 18 irregular particles and poly(styrene–divinylbenzene) membrane] for anionic analytes was first evaluated using a mixture of peptide standards to determine the overall sensitivity of this approach. These chromatographic preconcentrators provided an enhancement of sample loadings of up to 5 μl with good linear response and low n M concentration detection limits for most peptides investigated. The application of cPC–CZE–ES-MS is further demonstrated for extracts of O-deacylated LPS obtained from H. influenzae strain Eagan. In combination with novel enzymatic releasing methods using proteinase K, this technique provides unparalleled sensitivity and enabled the identification of LPS surface antigens from as little as five bacterial colonies.
ISSN:0021-9673
DOI:10.1016/S0021-9673(98)00341-0