This study investigated the effect of UV-B irradiation and the combinational effect with glucosamine caramel, fructosazine and riboflavin on the antimicrobial activities against Bacillus subtilis (ATCC 6633) and two strains of Escherichia coli (AW 1.7 and ATCC 25922). The quantum yield of fructosazine was two times less than that of tryptophan, indicating its ability to emit fluorescent light but less efficiently than tryptophan. UV-B treatment alone was efficient to achieve a bactericidal effect for both E. coli stains tested, however no effect was found for Bacillus subtilis for up to 80 mJ/cm2 UV-B. The combination of UV-B with photosensitizers fructosazine, glucosamine caramel and riboflavin enhanced the UV-B efficacy against E. coli strains at lower UV-B doses, while Bacillus subtilis ATCC 6633 was more resistant to the treatment combinations. High-performance liquid chromatography showed the production of different fructosazine reaction products occurred during irradiation, including the possible formation of endoperoxides.