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  • Development of a new quantitative method for detection of endotoxin by fluorescence labeling of 3-hydroxy fatty acid.

Development of a new quantitative method for detection of endotoxin by fluorescence labeling of 3-hydroxy fatty acid.

Advances in experimental medicine and biology (1990-01-01)
K Tanamoto
ABSTRACT

New quantitative method for the detection of minute amounts of endotoxin has been developed using 3-hydroxytetradecanoic acid as a chemical marker. After converting 3-hydroxytetradecanoic acid to methyl ester, it was coupled with a fluorescent probe, anthracene-9-carboxyl chloride, obtained by chlorization of 9-anthroic acid with oxalyl chloride. The resulting ester was isolated by HPLC on silica column. The purified product, methyl-3-0-(9-carboxy-anthracenyl) tetradecanoate (M/Z 462), was highly responsive to a fluorescence spectrophotometer, showing maximum emission with excitation wavelength at 257 nm and emission wavelength at 458 nm in dichloromethane, the limit of detection being as little as 10 f mol. Using this method it is currently possible to detect Salmonella abortus equi endotoxin in aqueous solution at a level of 100 pg.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
DL-β-Hydroxymyristic acid, ≥98%