The hydroxylation and acetylation of 0.5 mM sulphadimidine (SDD) was studied in primary cultures of hepatocytes from male and female rats, and from castrated male and sham operated male rats. In addition, SDD metabolism was investigated in hepatocytes from castrated male rats treated with testosterone, prior to liver cell isolation. In male rat hepatocytes a significantly higher hydroxylation activity was observed than in hepatocytes from female and castrated male rats. Acetylation activity was higher in females. Testosterone induced hydroxylation but did not affect acetylation. These results correlate well with data from previous in vivo studies, showing the relevance of this in vitro model.