The production of hydroxyl free radical (.OH) by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in the absence of H2O2 was investigated by electron spin resonance with the use of the spin-trapping agent 5,5-dimethyl-1-pyrroline-1-oxide (DMPO). .OH was detected as well as a nitrogen-centered free radical (.NR) on exposure of 1.25 mM MNNG solution (pH 6.0) containing 25 mM DMPO to visible light from a tungsten-halogen lamp at an intensity of 6.68 mW/cm2. Production of .OH depended on the concentration of MNNG and on the intensity of light exposure, indicating that .OH production resulted from the photolysis of MNNG solution. Manganese-superoxide dismutase and catalase did not inhibit the production of .OH, excluding the possibility that .OH might be produced from O2-. or H2O2. When the concentration of DMPO was increased from 25 mM to 125 mM, the yield of .OH decreased significantly while that of .NR increased over twice that at 25 mM. When the concentration of DMPO was further increased to over 200 mM, only .NR was detected and its yield was the same as that at 125 mM, which indicates that DMPO inhibited .OH production but not through the trapping of .NR. From these results we suggest that nitric oxide (NO) derived with .NR from the photolysis of MNNG resulted in .OH production which was inhibited by DMPO in aqueous solution.
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IUBMB Life
January 2025
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