Practical use of chemical probes for reactive oxygen species produced in biological systems by γ-irradiation

Abstract Application of chemical probes, for detection of reactive oxygen species (ROS), was tested during γ-irradiation. The ethanol/α-(4-pyridyl-1-oxide)- N - tert -butylnitrone (4-POBN) and 3,3′-diaminobenzidine (DAB) were structurally stable enough to detect OH and H 2 O 2 , increasingly generated by γ-irradiation up to 1000 Gy. Interestingly, the production rate of H 2 O 2 , but not OH, during γ-irradiation, was significantly different between in vitro systems of lettuce and spinach. These results suggest that 4-POBN and DAB could be utilized as a semi-quantitative probe to quantify OH and H 2 O 2 , produced by γ-irradiation up to 1000 Gy.