Arsenazo III forms 2:1 complexes with Ca and 1:1 complexes with Mg under physiological conditions. Estimates of the apparent dissociation constants.

Experiments to determine the apparent dissociation constants of the Ca and Mg complexes of arsenazo III clearly indicated that the predominant Ca complex contains one Ca ion and two dye molecules, although previous reports have either claimed or assumed 1:1 complexing. The evidence is based on the effects of varying [dye] as well as [Ca] and [Mg], and clear evidence for the formation of 1:1 complexes with Ca was obtained only at submicromolar [dye], whereas Mg formed 1:1 complexes exclusively. The implications of these findings with regard to the use of arsenazo III as an indicator of intracellular free [Ca] are discussed, with particular reference to its selectivity for Ca and the interference effects of other ions.