Solubility of endotoxins from Escherichia coli and Pseudomonas aeruginosa
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/264172
- author
- Eduard, W ; Westby, MH and Larsson, Lennart LU
- organization
- publishing date
- 2004
- 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 Inc.
- 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
- 2016-04-01 11:53:02
- date last changed
- 2022-01-26 19:37:16
@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}}, keywords = {{accuracy; endotoxins; solubility; Limulus amebocyte lysate test}}, language = {{eng}}, number = {{4}}, pages = {{375--377}}, publisher = {{John Wiley & Sons Inc.}}, 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}}, doi = {{10.1002/ajim.20089}}, volume = {{46}}, year = {{2004}}, }