Alkaline forms of the ferric bis(N-acetylated) heme undecapeptide of cytochrome c (N-ac-HUP) and some of its derivatives have been studied by electronic absorption and electron paramagnetic resonance spectroscopies. Surprisingly, even at pH >12, no evidence could be found for the formation of a hydroxyl ion adduct, in direct contrast to a previous report concerning ferric heme peptides encapsulated in detergent micelles (Mazumdar et al. Inorg. Chem. 1991, 30, 700-705). A spectroscopically determined pK(a) of approximately 9 is assigned to the deprotonation of the constituent histidine ligand of heme iron in N-ac-HUP. The present findings are not entirely in keeping with those of an earlier study concerning the properties of N-acetylated heme octapeptide (Wang et al. J. Biol. Chem. 1992, 35, 15310-15318), the differences observed being attributed to the buffering media employed in the two investigations. The implications of the current results in relation to a better understanding of the alkaline transitions observed in hemoglobins and myoglobins is considered.