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Solubility of endotoxins from Escherichia coli and Pseudomonas aeruginosa

Eduard, W; Westby, MH and Larsson, Lennart LU (2004) In American Journal of Industrial Medicine 46(4). p.375-377
Abstract
Background The Limulus amebocyte lysate (LAL) assay may underestimate endotoxins because only soluble endotoxins are determined. The solubility of endotoxins was, therefore, studied in two species of Gram-negative bacteria. Methods Cultures were grown in serum broth, cells were harvested by centrifugation and washed in physiological saline. Bacterial suspensions were either filtered through PTFE filters and air dried at room temperature, dried in polypropylene tubes at room temperature in a desiccator, or freeze dried. Samples were extracted with aqueous Tween 20. The soluble and insoluble fractions were dried, methanolysed, and hydroxy-fatty acid methyl esters were determined by gas chromatography (GC) as markers of endotoxins. Results... (More)
Background The Limulus amebocyte lysate (LAL) assay may underestimate endotoxins because only soluble endotoxins are determined. The solubility of endotoxins was, therefore, studied in two species of Gram-negative bacteria. Methods Cultures were grown in serum broth, cells were harvested by centrifugation and washed in physiological saline. Bacterial suspensions were either filtered through PTFE filters and air dried at room temperature, dried in polypropylene tubes at room temperature in a desiccator, or freeze dried. Samples were extracted with aqueous Tween 20. The soluble and insoluble fractions were dried, methanolysed, and hydroxy-fatty acid methyl esters were determined by gas chromatography (GC) as markers of endotoxins. Results The solubility of the endotoxins from Escherichia coli and Pseudomonas aeruginosa ranged from 9% to 83%. Species and drying conditions had substantial influence on the solubility. Conclusions Endotoxin exposure may be underestimated in environmental samples by the LAL test because a substantial fraction can be non-soluble and is not detected. (C) 2004 Wiley-Liss, Inc. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
accuracy, endotoxins, solubility, Limulus amebocyte lysate test
in
American Journal of Industrial Medicine
volume
46
issue
4
pages
375 - 377
publisher
John Wiley & Sons
external identifiers
  • pmid:15376280
  • wos:000224405300010
  • scopus:4944227493
ISSN
0271-3586
DOI
10.1002/ajim.20089
language
English
LU publication?
yes
id
89a31be4-77c2-4fb3-9107-9ec60f6fa1ea (old id 264172)
date added to LUP
2007-10-22 07:52:59
date last changed
2017-01-01 04:37:58
@article{89a31be4-77c2-4fb3-9107-9ec60f6fa1ea,
  abstract     = {Background The Limulus amebocyte lysate (LAL) assay may underestimate endotoxins because only soluble endotoxins are determined. The solubility of endotoxins was, therefore, studied in two species of Gram-negative bacteria. Methods Cultures were grown in serum broth, cells were harvested by centrifugation and washed in physiological saline. Bacterial suspensions were either filtered through PTFE filters and air dried at room temperature, dried in polypropylene tubes at room temperature in a desiccator, or freeze dried. Samples were extracted with aqueous Tween 20. The soluble and insoluble fractions were dried, methanolysed, and hydroxy-fatty acid methyl esters were determined by gas chromatography (GC) as markers of endotoxins. Results The solubility of the endotoxins from Escherichia coli and Pseudomonas aeruginosa ranged from 9% to 83%. Species and drying conditions had substantial influence on the solubility. Conclusions Endotoxin exposure may be underestimated in environmental samples by the LAL test because a substantial fraction can be non-soluble and is not detected. (C) 2004 Wiley-Liss, Inc.},
  author       = {Eduard, W and Westby, MH and Larsson, Lennart},
  issn         = {0271-3586},
  keyword      = {accuracy,endotoxins,solubility,Limulus amebocyte lysate test},
  language     = {eng},
  number       = {4},
  pages        = {375--377},
  publisher    = {John Wiley & Sons},
  series       = {American Journal of Industrial Medicine},
  title        = {Solubility of endotoxins from Escherichia coli and Pseudomonas aeruginosa},
  url          = {http://dx.doi.org/10.1002/ajim.20089},
  volume       = {46},
  year         = {2004},
}