Applicability of new spin trap agent, 2-diphenylphosphinoyl-2-methyl-3,4-dihydro-2H-pyrrole N-oxide, in biological system

Electron spin resonance using spin-trapping is a useful technique for detecting direct reactive oxygen species, such as superoxide (O2·-). However, the widely used spin trap 2,2-dimethyl-3,4-dihydro-2H-pyrrole N-oxide (DMPO) has several fundamental limitations in terms of half-life and stability. Recently, the new spin trap 2-diphenylphosphinoyl-2-methyl-3,4-dihydro-2H-pyrrole N-oxide (DPhPMPO) was developed by us. We evaluated the biological applicability of DPhPMPO to analyze O2·- in both cell-free and cellular systems. DPhPMPO had a larger rate constant for O2·- and formed more stable spin adducts for O2·- than DMPO in the xanthine/xanthine oxidase (X/XO) system. In the phorbol myristate acetate-activated neutrophil system, the detection potential of DPhPMPO for O2·- was significantly higher than that of DMPO (kDMPO = 13.95 M−1 s−1, kDPhPMPO = 42.4 M−1 s−1). These results indicated that DPhPMPO is a potentially good candidate for trapping O2·- in a biological system.