Solubility of endotoxins from Escherichia coli and Pseudomonas aeruginosa.
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.
Eduard
Wijnand
W
National Institute of Occupational Health, Oslo, Norway. Winjaud.Eduard@stami.no
Westby
Mette Haraldsen
MH
Larsson
Lennart
L
D001427:Bacterial Toxins / Q000032:analysis* / Q000302:isolation & purification; D002849:Chromatography, Gas; D004731:Endotoxins / Q000032:analysis* / Q000302:isolation & purification; D004926:Escherichia coli / Q000737:chemistry*; D005227:Fatty Acids / Q000737:chemistry; D011550:Pseudomonas aeruginosa / Q000737:chemistry*; D012995:Solubility
D001427:Bacterial Toxins; D004731:Endotoxins; D005227:Fatty Acids