Gaining an understanding of the glucose oxidation product (glyoxal) interaction with the metal–organic complex in a biological environment is pivotal to its usage. Thus, the glyoxal (Gyx) oxidation with Mo-oxime complex (MC) is studied with the spectrophotometric method, following a pseudo-phase approach. The result highlights the inclusion of hydrolysis, ion catalysis, the neutral primary salt effect, and radical generation as the essential factors in Gyx oxidation, with the exclusion of intermediate species formation. The hydrolysis of Gxy is observed to actively engage MC without charge inhibition as charge-neutral reacting species are involved, thus, implicating neutral primary salt effect where the variation of ionic strength of the system keeps the redox rate unchanged. The involvement of charge-neutral specie at the rate controlling step encouraged electrostatic attraction when Mg2+ ion additive is incorporated into the system, leading to ion catalysis. The generation of free radical from Gxy aids the emergence of formic acid. The zero intercept of Michealis-Menten type plot and the non-appreciable shift in the maximum absorption wavelength of the reaction system and MC rule-out the presence of intermediate species. The contribution of thermodynamic enthalpy is instrumental in the redox process, leading to the formic acid product. The inclusion of benzalkonium chloride (BZC) hastens the Gxy oxidation, which is supported by Raghavan and Srinivasan’s model.
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