Determination of ascorbic acid in pharmaceutical dosage forms and urine by means of an oscillatory reaction system using the pulse perturbation technique
A simple and reliable method for the determination of ascorbic acid (AA) is proposed and validated. It is based on potentiometric monitoring of the concentration perturbations of an oscillatory reaction system in a stable nonequilibrium stationary state close to the bifurcation point. The response of the Bray–Liebhafsky (BL) oscillatory reaction as a matrix, to the perturbation by different concentrations of AA, is followed by a Pt electrode. The linear relationship between maximal potential shift and the logarithm of the amount of AA is obtained between 0.01 and 1.0 μmol. The sensitivity of the proposed method (as the limit of detection) is 0.009 μmol and the method has excellent sample throughput (30 samples per hour). The procedure was used for AA determination in pharmaceutical formulations and urine. The results are in agreement with those obtained using the official method. Some aspects of the possible mechanism of AA action on the BL oscillating chemical system are discussed.
Determination of ascorbic acid based on a peroxidase oscillator reaction
Abstract This paper reports a new method for determination of ascorbic acid based on a peroxidase-oxidase (PO) biochemical oscillator reaction. The oscillation period changes linearly with concentration of ascorbic acid in the range 0.175–17.5 μM with a correlation coefficient of 0.9982 (n=6) based on oxygen measurement with an oxygen electrode. The results compared well with those obtained by an AOAC procedure.
Oscillations in the peroxidase-oxidase reaction: a comparison of different peroxidases.
The nonlinear behavior of the peroxidase-oxidase reaction was studied using structurally different peroxidases. For the first time sustained oscillations with peroxidases other than horseradish peroxidase in a single-enzyme system were observed. All peroxidases that showed significant oxidase activity were able to generate sustained oscillations. When adjusting the overall reaction rate, either of the two modifiers 2,4-dichlorophenol or Methylene blue could be omitted from the reaction. Due to the observation of different enzyme intermediates when using different peroxidases, we conclude that the mechanisms responsible for oscillatory kinetics may vary from one peroxidase to the other.