Open AccessA micro‐quantitative method for direct detection of bacterial endotoxin by fluorescence labelling of l‐ glycero ‐d‐ manno ‐heptose
Author(s) -
Haishima Yuji,
Tanamoto Kenichi
Publication year - 1993
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1993.tb06446.x
Subject(s) - fluorescence , chemistry , labelling , chromatography , analytical chemistry (journal) , biochemistry , physics , optics
A new method for analysis of bacterial endotoxin was developed using l‐ glycero ‐d‐ manno ‐heptose (ld‐Hep) as a chemical marker. Authentic ld‐Hep was coupled with a fluorescence probe, 2‐aminopyridine, by pyridylamination. The product, pyridylaminated ld‐Hep, was separated by high performance liquid chromatography and the fluorescence intensity was measured by a highly sensitive detector system by excitation at 300 nm and emission at 368 nm in 0.7 M H 3 BO 3 ‐KOH buffer (pH 9.0)/CH 3 CN (9:1, v/v). The fluorescence intensity of ld‐Hep increased in a dose‐dependent manner over the range of 30 pg to 100 μg tested. As little as 400 pg of Salmonella abortus equi endotoxin could be quantitatively measured by this method.