Localized excess protons are created with an open-circuit water electrolysis process using a pair of anode and cathode electrodes for a special excess proton production and proton-utilization system to treat a substrate material plate/film by forming and using an excess protons-substrate-hydroxyl anions capacitor-like system. The technology enables protonation and/or proton-driven oxidation of plate/film and/or membrane materials in a pure water environment. The present invention represents a remarkable clean green chemistry technology that does not require the use of any conventional acid chemicals including nitric and sulfuric acids for the said industrial applications. The application of localized excess protons provides a special energy recycling and renewing technology function to extract latent heat including molecular thermal motion energy at ambient temperature for generating local proton motive force (equivalent to Gibbs free energy) to do useful work such as driving ATP synthesis and proton-driven oxidation of certain substrate metal atoms.

This disclosure relates to methods for determining pH and also calcium (Ca.sup.2+) concentration or chloride (Cl.sup.) concentration in biological samples. More particularly, this disclosure relates to methods capable of simultaneously determining pH and Ca.sup.2+ concentration, or pH and Cl.sup. concentration using nucleic acid complexes.