SOFT 2004 Meeting Abstracts - Society of Forensic Toxicologists

A45
FAST QUANTIFICATION OF ETHANOL IN WHOLE BLOOD SPECIMENS BY THE
ENZYME ALCOHOL DEHYDROGENASE METHOD; OPTIMIZA TION BY
EXPERIMENTAL DESIGN
Lena Kristoffersen·· l , Bjorn Skuterud 1 , Bente R. Larssen 1 , Svetlana Skurtveit 2 , Anne Smith­
Kielland l
lNorwegian Institute of Public Health, Division of Forensic Toxicology and Drug Abuse, P.O. Box
4404 Nydalen, N-0403, Oslo, Norway; 2Norwegian Institute of Public Health, Division of
Epidemiology, P.O. Box 4404 Nydalen, N-0403, Oslo, Norway
A sensitive, fast, simple and high throughput enzymatic method for the quantification of ethanol in
whole blood (blood) on Hitachi 917 is presented. Alcohol dehydrogenase (ADH) oxidizes ethanol
to acetaldehyde using the coenzyme nicotinamide adenine dinucleotide (NAD), which is
concurrently reduced to form NADH. Method development was performed with the aid of factorial
design, varying pH and concentrations ofNAD+ and ADH. The linear range increased and reaction
end point decreased with increasing NAD+ concentration and pH. The method was linear in the
concentration range 0.0024-0.4220 g/d!. The limit of detection (LOD) and limit of quantification
(LOQ) were 0.0007 g/dl and 0.0024 gldl, respectively. Relative standard deviations (RSD) for the
repeatability and within laboratory reproducibility were in the ranges 0.7-5.7 % and 1.6-8.9 %,
respectively. The correlation coefficient when compared to head space gas chromatography flame
ionization detection (HS-GC-FID) methods was 0.9903. Analysis of authentic positive blood
specimens gave results that were slightly lower than those of the reference method.
Key words: Ethanol, ADH, Quantitation
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