Incubation of human colonic contents with various ethanol concentrations (2.75-44 mM) in vitro at 37 degrees C resulted in significant accumulation of acetaldehyde--a toxic and highly reactive compound. At pH 9.6, all samples produced notable acetaldehyde concentrations (58 (13) microM; mean (SEM)) even from the lowest (2.75 mM) ethanol concentration, and the production of acetaldehyde increased lin-early with rising ethanol concentration (r = 0.97; p < 0.005), reaching a peak concentration of 238 (37) microM at 44 mM ethanol. The formation of acetaldehyde took place rapidly, as almost 50% of acetaldehyde formed during the total eight hour incubation was detectable after one hour, and 75% of the total after four hours. Maximal acetaldehyde production from 22 mM ethanol occurred at pH 9.6 (160 (35) microM) but appreciable concentrations were also seen at pH 7.4 (110 (38) microM) and pH 6.0 (63 (19) microM). At pH 4.0, by contrast, acetaldehyde formation was negligible (17 (5) microM). 4-Methylpyrazole, a potent inhibitor of alcohol dehydrogenase, showed a decreasing effect on acetaldehyde production in vitro but first at a concentration of 100 mM. Considerable acetaldehyde production by human colonic bacteria--if it occurs also in vivo--could constitute a risk factor for rectal cancer in heavy drinkers and also provide a pathogenetic mechanism for alcohol induced diarrhoea.