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

Publication ,  Journal Article
Li, J; Thibault, P; Martin, A; Richards, JC; Wakarchuk, WW; van der Wilp, W
Published in: J Chromatogr A
August 21, 1998

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., C18 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 microliters with good linear response and low nM 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.

Duke Scholars

Published In

J Chromatogr A

DOI

ISSN

0021-9673

Publication Date

August 21, 1998

Volume

817

Issue

1-2

Start / End Page

325 / 336

Location

Netherlands

Related Subject Headings

  • Sensitivity and Specificity
  • Molecular Sequence Data
  • Mass Spectrometry
  • Lipopolysaccharides
  • Haemophilus influenzae
  • Electrophoresis, Capillary
  • Carbohydrate Sequence
  • Analytical Chemistry
  • Amino Acid Sequence
  • 10 Technology
 

Citation

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Li, J., Thibault, P., Martin, A., Richards, J. C., Wakarchuk, W. W., & van der Wilp, W. (1998). Development of an on-line preconcentration method for the analysis of pathogenic lipopolysaccharides using capillary electrophoresis-electrospray mass spectrometry. Application to small colony isolates. J Chromatogr A, 817(1–2), 325–336. https://doi.org/10.1016/s0021-9673(98)00341-0
Li, J., P. Thibault, A. Martin, J. C. Richards, W. W. Wakarchuk, and W. van der Wilp. “Development of an on-line preconcentration method for the analysis of pathogenic lipopolysaccharides using capillary electrophoresis-electrospray mass spectrometry. Application to small colony isolates.J Chromatogr A 817, no. 1–2 (August 21, 1998): 325–36. https://doi.org/10.1016/s0021-9673(98)00341-0.
Journal cover image

Published In

J Chromatogr A

DOI

ISSN

0021-9673

Publication Date

August 21, 1998

Volume

817

Issue

1-2

Start / End Page

325 / 336

Location

Netherlands

Related Subject Headings

  • Sensitivity and Specificity
  • Molecular Sequence Data
  • Mass Spectrometry
  • Lipopolysaccharides
  • Haemophilus influenzae
  • Electrophoresis, Capillary
  • Carbohydrate Sequence
  • Analytical Chemistry
  • Amino Acid Sequence
  • 10 Technology