Flavins are a unique class of compounds that combine the features of singlet oxygen generators and redox-dependent fluorophores. From a broad family of flavin derivatives, deazaalloxazines are significantly underdeveloped from the point of view of photophysical properties. Herein, we report photophysics of 5-deazaalloxazine (1a) in water, acetonitrile, and some other solvents. In particular, triplet excited states of 1a in water and in acetonitrile were investigated using ultraviolet-visible (UV-Vis) transient absorption spectroscopy. The measured triplet lifetimes for 1a were all on the microsecond time scale (≈ 60 μs) in deoxygenated solutions. The quantum yield of S1 → T1 intersystem crossing for 1a in water was 0.43 based on T1 energy transfer from 1a to indicaxanthin (5) acting as acceptor and on comparative actinometric measurements using benzophenone (6). 1a was an efficient photosensitizer for singlet oxygen in aerated solutions, with quantum yields of singlet oxygen in methanol of about 0.76, compared to acetonitrile ~ 0.74, dichloromethane ~ 0.64 and 1,2-dichloroethane ~ 0.54. Significantly lower singlet oxygen quantum yields were obtained in water and deuterated water (ФΔ ~ 0.42 and 0.44, respectively). Human red blood cells (RBC) were used as a cell model to study the antioxidant capacity in vitro and cytotoxic activity of 1a. Fluorescence-lifetime imaging microscopy (FLIM) data were analyzed by fluorescence lifetime parameters and distribution for different parts of the emission spectrum. Comparison of multidimensional fluorescent properties of RBC under physiological-like and oxidative-stress conditions in the presence and absence of 1a suggests its dual activity as probe and singlet-oxygen generator and opens up a pathway for using FLIM to analyze complex intracellular behavior of flavin-like compounds. These new data on structure-property relationship contribute to the body of information required for a rational design of flavin-based tools for future biological and biochemical applications.

Department of Chemistry and Pharmacy Faculty of Sciences and Technology and Centre for Electronics Optoelectronics and Telecommunications University of the Algarve 8005 139 Faro Portugal

Department of Organic Chemistry University of Chemistry and Technology Prague Technicka 5 Prague 6 16628 Prague Czech Republic

Faculty of Biology Adam Mickiewicz University Uniwersytetu Poznańskiego 6 61 614 Poznań Poland

Faculty of Chemistry Adam Mickiewicz University Uniwersytetu Poznańskiego 8 61 614 Poznań Poland

Faculty of Physics Adam Mickiewicz University Uniwersytetu Poznańskiego 2 61 614 Poznań Poland

Institute of Bioorganic Chemistry Polish Academy of Sciences Noskowskiego 12 14 61 704 Poznań Poland

Poznań University of Economics and Business Al Niepodległości 10 61 875 Poznań Poland

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A facile three-component route to powerful 5-aryldeazaalloxazine photocatalysts

Tetramethylalloxazines as efficient singlet oxygen photosensitizers and potential redox-sensitive agents