Investigation of free radical metabolism using microdialysis and HPLC7ESR/MS

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In order to detect 4-POBN/phenyl radical adducts obtained from microdialysis, it is essential to develop a high sensitive detection system. We have developed an HPLC system which uses both ultraviolet detector and electrochemical detector. The electrochemical detector allowed us to detect both reduced and radical forms of 4-POBN/phenyl radical adduct. Futhermore, concentration of the 4-POBN/phenyl radical adduct can be estimated by analysis of peak area of elctrochemical detector.Ultraviolet absorption spectra of reduced, radical, and oxidized forms of 4-POBN/phenyl radical adducts were measured. The lambdamax and epsilonmax obtained are as follows ; reduced from [lambdamax : 214 nm, epsilonmax : 2.34 * 10^4 (cm・l/mol) ; lambdamax : 266 nm, epsilonmax : 2.91 * 10^4 (cm・l/mol)], radical form [lambdamax : 215 nm, epsilonmax : 1.88 * 10^4 (cm・l/mol) ; lambdamax : 265 nm, epsilonmax : 1.74 * 10^4(cm・l/mol)], and oxidized form [lambdamax : 215 nm, epsilonmax : 1.85 * 10^4 (cm・l/mol) ; lambdamax : 243 nm, epsilonmax : 1.35 * 10^4 (cm・l/mol) ; lambdamax : 263 nm, epsilonmax : 1.40 * 10^4 (cm・/mol) ; lambdamax : 330 nm, epsilonmax : 1.76 * 10^4 (cm・l/mol)].