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68 Z. Rechtsmedizin 1 (1971)

handle is hein.journals/injlegame68 and id is 1 raw text is: Z. Rechtsmedizin 68, 1-6 (1971) © by Springer-Verlag 1971 Originalarbeiten -Original Papers Influence of Pyrazole on Ethanol Determination with the automated ADH Method ULF RYDBERG, JAN BUIJTEN, LENA DAHLGREN and JAN DOCK Department of Alcohol Research, Karolinska Institutet, and the Alcohol Clinic, Karolinska Sjukhuset, Stockholm (Sweden) Received May 15, 1970 Summary. It has been demonstrated that the contamination of blood samples with pyrazole will disturb automated enzymatic determinations of ethanol by methods involving the use of YADH, if the sample is not distilled. Determination of ethanol by gas liquid chromatography, the Widmark method or by the ultramicro destillation enzymatic method for ethanol analysis (AA-UMDE) according to Buijten were not disturbed by the presence of pyrazole. Contamination of samples with 4-iodo-pyrazole has not been found to disturb ethanol determinations by any method. Key- Words: Enzymatic methods for ethanol-Pyrazole, Influence on. Zusacnrnenfassung. Es wurde die Kontamination der Blutprobe mit Pyrazol in wallrigen Losung als Salbe verarbeitet sowie als 4-Iodo-Pyrazol-Losung appliziert iiberpriift. Bei der An- wendung der Hefe-ADH zur automatisierten enzymatischen Blutalkoholbestimmung ohne Destillationsverfahren treten Storungen ein. Die Anwendung der Gas-chromatographischen Methode, des Widmark-Verfahrens sowie der enzymatischen Ujltramikromethode nach Buijten verursacht keine Beeinflussung durch Pyrazol-Beimischung. Die Bestimmungen des Blut- alkohols mittels dieser Methoden warden nach Kontamination mit 4-Iodo-Pyrazol nicht beeinfluBt. Auf die mogliche forensische Bedeutung wird hingewiesen. Enzymatic methods involving an oxidation of NAD, catalyzed by alcohol dehydrogenase, and the absorption of formed NADH measured photometrically at a wavelength of 340 or 366 1m, belong to the most utilized methods for determination of ethanol. Various modifications have been described and the speci- ficity of the ADH methods has been critically discussed. Also a number of other substances than ethanol react with LADH (horse liver ADH) and YADH (yeast ADH). The oxidation of a number of higher aliphatic alcohols, esters, various steroids, retinol etc., will also be catalyzed by ADH (Theorell, 1969). The interference by such substances with enzymatic methods for ethanol determination has been demonstrated (Dotzauer et al., 1952; Hilgermann, 1970; Weinig et al., 1963). Theorell and Yonetani (1963) published the first experimental work on in vitro studies of pyrazole and analogs as inhibitors of LADH. A large number of com- pounds has been investigated, as published by Lester et at. (1968) and by Theorell et al. (1969). Pyrazole, 4-bromo-pyrazole, 4-iodo-pyrazole and 4-methyl-pyrazole were found as the most potent inhibitors yet known. The inhibition of ethanol metabolism ire vivo in experimental animals by pyrazole and analogs has also been investigated (Goldberg and Rydberg, 1969; 1 Z. Rechtsmedizin, Bd. 68

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