Photolysis of thiochrome in aqueous solution: A kinetic study

The photolysis of thiochrome (THC), an oxidation product of thiamine (vitamin B₁) (THE), used for its fluorimetric assay, has been studied in the pH range 7.0-12.0. THC undergoes photooxidation to oxodihydrothiochrome (ODTHC) which is oxidized to a non-fluorescent compound (OP1) on UV irradiation. The kinetics of the consecutive first-order reactions: THC→k₁ODTHC→k₂OP1, has been evaluated and the values of first-order rate constants, k₁ (0.58-4.20 × 10^-5, s^-1) and k₂ (0.05-2.03 × 10^-5, s^-1), at pH 7.0-12.0 have been determined. The rates of degradation of THC and ODTHC are enhanced with pH and the second-order rate constants k₁' and k₂' for the OH^- ion-catalyzed reaction are in the range of 0.002-58.3 M^-1 s^-1. The quantum yields of the photolysis of THC and ODTHC in the pH range 7.0-12.0 have been determined. THC, ODTHC and OP1 have been identified by chromatographic, spectrometric and fluorimetric methods. THC and ODTHC have similar fluorescence characteristics and emit at 450 and 445 nm, respectively. THC, ODTHC and OP1 with distinct absorption maxima (370, 344 and 290 nm, respectively) have been determined by a newly developed and validated multicomponent spectrometric method during the photolysis reactions. The on-line formation of THC by the photooxidation of THE may lead to the degradation of THC and give erroneous results in the fluorimetric assay of THE. A scheme for the photolysis reactions of THC in aqueous solution is presented.