The electrochemical redox behavior of the polynuclear mixed valence ruthenium oxide cyanometallate complexes (mvRuOxÀMCN, M Fe, Cr, Ni, Cu, Ru and Pt) have been systematically studied in this report by using three redox sensitive organic probes of glucose, ethanol and formaldehyde. The results were interpreted by the well-established ruthenium oxide and Prussian blue chemistry. The mvRuOxÀMCN, under the category of Ru-based Prussian blue analogue, was found to possess superior electrocatalytic activity than either ruthenium oxide or Prussian blue in acidic mediums. The electrogenerated oxy/hydroxy-Ru VII state (at 1.1 V vs. Ag/AgCl) was unusually stabilized in the mvRuOxÀMCN matrix without any disproportion reaction in acidic environments. In contrast to those of earlier studies, possible structure in terms of the ÀRu III/II ÀNCÀMÀand ÀRu III/II ÀOÀRu VII/VI Àsites was proposed here. Enzymeless analytical detection of glucose in acidic conditions was first time demonstrated with sensitivity comparable to that of ruthenium oxide-based electrodes in alkaline solutions.