Reaction Mechanism Spectroscopy Studies of Protein and Orange II

Abstract

In this paper, the interaction of Orange II and Human Serum Albumin was studied by fluorescence spectroscopy under simulated physiological conditions. The affection of pigment and serum albumin may cause changes in the microenvironment. Fluorescence spectroscopy using synchronous fluorescence can provide a reference for the micro-environment changes in serum albumin. By calculating quenching constantsat different temperatures, its quenching mechanism which is static quenching was determined. The Orange?and HSA binding constants and binding sites were also calculated and the electrostatic force of Orange? with HSA was determined according to the thermodynamic formula. According to the Forster theory, the energy transfer efficiency and biding distance of orange? between HSA were measured and the energytransfer quenching mechanism by the energy transfer was elaborated. Further application of UV absorption spectroscopy to study the mechanism of action and the conclusion of orange? fluorescence spectra are consistent with the HSA.