The enantiomers of 3-O-methyldobutamine, a metabolite of dobutamine, were evaluated for their alpha- and beta-adrenoceptor mediated effects in vitro in a variety of isolated organs and in radioligand binding studies. Neither enantiomer of 3-O-methyldobutamine possessed alpha 1-adrenoceptor agonist activity in isolated guinea pig aorta. However, both enantiomers of 3-O-methyldobutamine were competitive alpha 1-adrenoceptor antagonists, with the (+)-enantiomer being approximately 10-fold more potent than the (-)-enantiomer as assessed either in guinea pig aorta or by displacement of 3H-prazosin binding from alpha 1-adrenoceptors in rat cerebral cortex. The alpha 1-adrenoceptor blocking activity of (+)-3-O-methyldobutamine was relatively potent and corresponded to a pA2 of 7.33 in guinea pig aorta and a -log Ki of 7.72 in radioligand binding studies. Neither enantiomer of 3-O-methyldobutamine possessed alpha 2-adrenoceptor agonist activity in field-stimulated guinea pig ileum. Although (+)-3-O-methyldobutamine weakly inhibited the twitch response in field-stimulated guinea pig ileum, the response was not blocked by the selective alpha 2-adrenoceptor antagonist, yohimbine, and was found to result from weak anticholinergic activity (pA2 = 5.06). Neither enantiomer of 3-O-methyldobutamine possessed beta 1-adrenoceptor agonist activity in guinea pig atria, however the (+)-enantiomer was a weak noncompetitive antagonist at beta 1-adrenoceptors. In contrast, both enantiomers of 3-O-methyldobutamine were weak beta 2-adrenoceptor agonists in rat uterus, however these weak effects were not highly stereoselective, which was also confirmed in radioligand binding studies.(ABSTRACT TRUNCATED AT 250 WORDS)