Fluorimetry Assay of Hydroxyl Radical by Fenton-Like Reaction-Aromatic Hydroxylation System

In this study, RP-HPLC with fluorescence detection(excitation 277 nm and emission 306 nm)was used to detect hydroxyl radical generated by Fenton like reaction system participatated by seven transition metal-ions,using a fluorescence probe,L-phenylalanine,which could be converted to highly fluorescent L-tyrosine in the generation of ·OH the variety of fluorescence intensity reflected the generation of ·OH.The experimental results indicated that ability of Cr(Ⅲ),Fe(Ⅱ),Co(Ⅱ),Cu(Ⅱ),Pb(Ⅱ) to produce ·OH is relatively.strong,but the ability of Cd(Ⅱ) and Ni(Ⅱ) is poor. The intensity of fluorescence in the systems could be inhibited obviously by mannite and desferrioxamine(DFO),which proved ·OH was cleard. We used the UV spectrometer and got the same results. Aromatic hydroxylation was more effective and reliable. So this sensitive method can be applied to study the hydroxyl radical and hydroxyl radical scavenger.