A quantitative microanalysis of bacterial endotoxin using [3H]-labeled L-glycero-D-mannoheptitol as a marker.
Quantitative microanalysis of bacterial endotoxin was performed using [3H]-labeled L-glycero-D-mannoheptitol (LD-Heptitol) as a marker. Several different amounts of authentic L-glycero-D-mannoheptose (LD-Heptose) were reduced with 20 micrograms of cold NaBH4 containing 2 micrograms of NaB3H4 (40 Ci/mmol) in 20 microliters of 1 mM NaOH at 4 C for 48 hr. The product, [1-3H]-labeled LD-Heptitol, has high specific activity, and was purified by HPLC and detected using a liquid-scintillation counter. As little as 50 pg of LD-Heptose was detectable, and the radioactivity increased dose-dependently in the 100 pg to 80 ng range tested. More than 2 ng of Salmonella abortus equi endotoxin could be accurately determined by this method. It is possible to detect 50 pg of endotoxin by this method, if 100% hot material (NaB3H4) is used for [3H]-labeling.
Haishima
Y
Y
Department of Microbiology, National Institute of Hygienic Sciences, Tokyo, Japan.
Tanamoto
K
K
D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004731:Endotoxins / Q000032:analysis*; D006539:Heptoses / Q000737:chemistry*; D012475:Salmonella / Q000737:chemistry*; D014316:Tritium
D004731:Endotoxins; D006539:Heptoses; D014316:Tritium; C026312:glycero-manno-heptose; C025642:perseitol