In this study, the effect of light activated agent, methyl (pyro) pheophorbide-a, which bears non-aromatic cyclic compound, excited with red light from a LED on the viability of S. aureus, S. epidermidis, and E. coli was investigated. All species were susceptible to killing by photosensitization and photodynamic effect was dependent on both the chemical structure and concentration. However, E. coli was not susceptible to concentrations used to obtain a significant kill with the Gram-positive bacteria upon irradiation. To more closely mimic the conditions of wounds, photodynamic therapy was carried out on S. aureus, which is the most important organism that can cause a range of mild to severe infections in skin and burn wounds, in the presence of human blood plasma and human serum albumin, representing a wound fluid model. Results indicate that microorganisms could be successfully photoinactivated by tropolone methyl (pyro) pheophorbide-a derivatives when suspended in phosphate buffered saline. However, changing the medium into 4.5% and 7% HSA/PBS solutions reduced the effectiveness of lethal photosensitization of bacteria. The same results were obtained with human blood plasma. Also, the mechanism of bacterial cell inactivation by a sensitizer and light was studied with reactive oxygen species scavengers. Further evidence of the involvement of singlet oxygen is provided by the protective effect of the singlet oxygen scavenger, sodium azide.