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The aim of the present study was to consider bilirubin (BRB) and riboflavin (RFL) mutual interaction in methanol solution under continuous UV-A and UV-B irradiation regime. Continuous irradiations of samples were performed in a cylindrical photochemical reactor “Rayonet”, with 10 symmetrically placed lamps having the emission maximum at 300 nm (UV-B) and 350 nm (UV-A). The rate of BRB and RFL photodegradation along with simultaneous products formation, as a function of UV exposure time, was followed by combining UV-VIS absorption measurements with RP-HPLC analysis. The compounds were separated by gradient elution with mobile phase A (formic acid, 0.1% water solution) and B (formic acid, 0.1% methanol solution). According to the results obtained, BRB degradation in the absence of RFL was almost 22 times and 9 times slower in comparison to its degradation observed in BRB-RFL mixture under continuous UV-A and UV-B irradiation, respectively. Moreover, BRB degradation in BRB-RFL mixture under anaerobic conditions was almost 24 times and 16 times slower in comparison to the degradation in aerobic conditions under UV-A and UV-B light, respectively. The latter observation suggests that presence of ROS species contributes to UV-induced BRB degradation. These experiments provide the indirect proof of BRB acting as Type II sensitizer because of the fact that 1O2 produced by RFL mediates BRB irreversible degradation giving rise to dipyrrole methanol adducts as typical products obtained via Type II mechanism.

BILIRUBIN-RIBOFLAVIN MUTUAL INTERACTION IN METHANOL UNDER CONTINUOUS UV IRRADIATION REGIME