Ethyl acetate extraction of spin-trapped free radicals: a reevaluation.

We have suggested the use of ethyl acetate for extraction of hydroxyl or superoxide radical adducts of the spin trap phenyl N-tert-butyl nitrone (PBM). The technique produced EPR spectra with narrow line widths, the radical adducts were more stable, and there were sufficiently large differences between the isotropic nitrogen hyperfine coupling constant (alpha N) and the beta hydrogen coupling constant (alpha H beta) for both the hydroxyl and superoxide radical adducts to allow their simultaneous quantitation in mixtures. However, Kalyanaraman, Mottley, and Mason have suggested that our assignments of alpha N and alpha H beta were incorrect and that extraction of spin-trapped adducts into ethyl acetate is not as useful as we had proposed. This paper demonstrates that their objections are unfounded and are based on a computational error that they made when they attempted to calculate the hyperfine splittings in their spectra.