Determination of the hydroxyl radical by its adduct formation with phenol and liquid chromatography/electrochemical detection.
An HPLC-ECD method is described for the indirect determination of the hydroxyl (OH) radical. Fenton's reaction is used to produce OH, which simultaneously attacks phenols (phenol or pyrocatechol) to form the adducts, pyrocatechol or pyrogallic acid. Thus, [OH] quantification is based on the separation and detection of pyrogallic acid and/or pyrocatechol by an isocratic HPLC-ECD method. The quantification of OH is also performed alternatively by a chronoamperometric detection in an electrochemical cell, where simultaneously formed Fe(III) (Fenton's reaction) combines [Fe(II)(CN)(6)](4-) to produce the Prussian blue (PB) molecules (Fe(4)(III)[Fe(II)(CN)(6)](3)). Newly formed PB molecules are then immediately converted to colorless Everitts salt (K(4)Fe(4)(II)[Fe(II)(CN)(6)](3)) with the reduction of the high-spin Fe(III) to Fe(II) at the surface of a glassy carbon electrode at +0.150V (versus Ag/AgCl). The calculated concentration of OH during incubation (0.626ppm) can be detected with negative errors by the HPLC-ECD (0.595 and 0.615ppm with the errors -5.2 and -1.8%, respectively) and by the chronoamperometric method (0.552 and 0.607ppm with the errors -11.8 and -3.0%, respectively). For the comparison of the two sets of data, HPLC-ECD method is much more promising.
Kilinc
Emrah
E
Faculty of Pharmacy, Department of Analytical Chemistry, University of Ege, 35100 Bornova, Izmir, Turkey.