Simultaneous detection of NO and ROS by ESR in biological systems.

A large body of evidence shows that the generation of nitric oxide (NO) and reactive oxygen species (ROS) and the rate of ROS/NO play an important role in the biological system. We developed a method to simultaneously detect NO free radical and ROS in biological systems using ERS spin trapping technique. The adduct (DETC)2-Fe2+-NO and N-tert- butyl-alpha-phenylnitrone (PBN)-ROS in biological systems can be extracted by organic solvent and then measured on an electron spin resonance (ESR) spectrometer at room temperature because the g = 2.035 of (DETC)2-Fe2+-NO is different from that of PBN-ROS (g = 2.005) and their ESR signals can be separated clearly. Using this method, we measured the production of NO and ROS in plant and animal systems.