Production of formaldehyde and acetone by hydroxyl-radical generating systems during the metabolism of tertiary butyl alcohol.
t-Butyl alcohol is not a substrate for alcohol dehydrogenase or for the peroxidatic activity of catalase and, therefore, it is used frequently as an example of a non-metabolizable alcohol. t-Butyl alcohol is, however, a scavenger of the hydroxyl radical. The current report demonstrates that t-butyl alcohol can be oxidized to formaldehyde plus acetone by hydroxyl radicals generated from four different systems. The systems studied were: (a) two chemical systems, namely, the iron catalyzed oxidation of ascorbic acid and the Fenton reaction between H2O2 and iron; (b) an enzymatic system, the coupled oxidation of xanthine by xanthine oxidase; and (c) a membrane-bound system, NADPH-dependent microsomal electron transfer. The oxidation of t-butyl alcohol appeared to be mediated by hydroxyl radicals, or by a species with the oxidizing power of the hydroxyl radical, because the production of formaldehyde plus acetone was (a) inhibited by competing scavengers of the hydroxyl radical; (b) stimulated by the addition of iron-EDTA; and (c) inhibited by catalase. The last observation suggests that H2O2 served as the precursor of the hydroxyl radical in all three systems. A possible mechanism is hydrogen abstraction to form the alkoxyl radical [CH3)3-C-O.), spontaneous fission of the alkoxyl radical to produce acetone and the methyl radical (CH3.), interaction of the methyl radical with O2 to form the methyl peroxy radical (CH300.), and decomposition of the later to formaldehyde. These results extend the alcohol oxidizing capacity of the microsomal alcohol oxidizing system to a tertiary alcohol. Since t-butyl alcohol is not a substrate for alcohol dehydrogenase or catalase, the ability of microsomes to oxidize t-butyl alcohol lends further support for a role for hydroxyl radicals in the microsomal alcohol oxidation system. In view of the production of formaldehyde, and the reactivity as well as further metabolism of this aldehyde, caution should be used in interpreting experiments in which t-butyl alcohol is used as a presumed "non-metabolizable" alcohol. t-Butyl alcohol may be a valuable probe for the detection of hydroxyl radicals in intact cells and in vivo.
D000096:Acetone / Q000378:metabolism*; D000818:Animals; D001205:Ascorbic Acid; D000440:Butanols / Q000378:metabolism*; D055598:Chemical Phenomena; D002621:Chemistry; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004492:Edetic Acid; D005557:Formaldehyde / Q000378:metabolism*; D005609:Free Radicals; D006861:Hydrogen Peroxide; D006878:Hydroxides / Q000378:metabolism*; D017665:Hydroxyl Radical; D066298:In Vitro Techniques; D007501:Iron; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D008956:Models, Chemical; D010084:Oxidation-Reduction; D051381:Rats; D011919:Rats, Inbred Strains; D019820:Xanthine; D014969:Xanthine Oxidase / Q000378:metabolism; D014970:Xanthines / Q000378:metabolism; D020002:tert-Butyl Alcohol
D000440:Butanols; D005609:Free Radicals; D006878:Hydroxides; D014970:Xanthines; D000096:Acetone; D019820:Xanthine; D005557:Formaldehyde; D017665:Hydroxyl Radical; D003577:Cytochrome P-450 Enzyme System; D004492:Edetic Acid; D006861:Hydrogen Peroxide; D007501:Iron; D014969:Xanthine Oxidase; D020002:tert-Butyl Alcohol; D001205:Ascorbic Acid