Silica sol-gel immobilized amperometric enzyme electrode for peroxide determination in the organic phase

Abstract

Abstract An amperometric enzyme electrode for peroxide determination in non-aqueous media has been constructed by the silica sol-gel immobilization of horseradish peroxidase (HRP) on a carbon paste electrode. Peroxides have been detected in the presence of ferrocene as the mediator in the organic phase to transfer electrons between the electrode surface and peroxides. The fabrication procedure of the enzyme electrode based on a sol-gel thin film derived from tetramethoxysilane has been optimized. The effect of operational parameters, such as different organic solvents, the amount of aqueous buffer in the organic solvent, mediator concentration and holding potential have been explored for the optimum analytical performance of the electrode. The electrode prepared using this immobilization technique has an excellent stability in some organic solvents, including methanol, acetonitrile and 2-propanol. The trend in the sensitivity of different peroxides is as follows: H 2 O 2 >lauroyl peroxide>butanone peroxide>tertbutyl-hydroperoxide. The apparent Michaelis–Menten constants ( K m app ) of the electrode in methanol for the above peroxides have been calculated. Applicability of the enzyme electrode for the measurement of hydrogen peroxide in real samples has been demonstrated. The results obtained by this electrode corroborate very well with the classical iodometric titration method for the determination of hydrogen peroxide.