beta-Funaltrexamine (beta-FNA) potently competed with the binding of a series of radiolabeled opiates and opioid peptides in standard binding assays with IC50 values under 10 nM. In addition, higher concentrations of beta-FNA produced an irreversible inhibition of binding which was relatively selective for mu receptors; delta binding was not affected much. The production of irreversible inhibition of [3H]dihydromorphine binding required concentrations of beta-FNA over 10-fold higher than beta-FNA concentrations needed in standard competition studies. Both mu 1 and mu 2 sites were irreversibly inhibited by beta-FNA, but mu 1 sites were more sensitive. The reversible and irreversible inhibition in these in vitro binding assays by beta-FNA were quite similar to naloxonazine. However, the activity of beta-FNA in the guinea-pig ileum suggests that it may not distinguish between mu 1 and mu 2 receptors as effectively as naloxonazine in bioassays and in vivo.