A new electrochemical method for the determination of microamounts of hydrogen peroxide utilizing o-dianisidine (ODA) as substrate and hemoglobin (Hb) as catalyst is described in this paper. Hb can be used as mimetic peroxidase and it can catalyse the reduction of hydrogen peroxide with the subsequent oxidation of ODA. The oxidative reaction product is an azo compound, which is an electroactive substance and has a sensitive second-order derivative polarographic reductive peak at the potential of -0.58 V (vs. SCE) in pH 80 Britton-Robinson (B-R) buffer solution. The conditions of Hb-catalytic reaction and polarographic detection of the reaction product were carefully studied. By using this polarographic peak and under optimal conditions, the calibration curve for the H₂O₂ was constructed in the linear range of 2.0 x 10^-7 ∼ 10 x 10^-4 mol/l with the detection limit of 5.0 x 10^-8 mol/l. This method can also be used to the determination of Hb content in the range of 20 x 10^-9 ∼ 30 x 10^-7 mol/l with a detection limit of 10 x 10^-9 mol/l. The proposed method was further applied to the determination of the content of H₂O₂ in fresh rainwater with satisfactory results. The catalytic reaction mechanism and the electrode reductive process of the reaction product were carefully studied.

An electrochemical method for the determination of lysozyme (LYS) based on its interaction with alizarin red S (ARS) was established by linear sweep voltammetry in this paper. The electrochemical behaviour of ARS with LYS was investigated on a dropping mercury working electrode in 0.2 mol/L pH 4.8 Britton-Robinson (B-R) buffer solution. ARS showed a sensitive second order derivative linear sweep voltammetric reductive peak at -0.42 V (vs SCE). After the addition of LYS, the reductive peak current of ARS decreased without the shift of the reductive peak potential and no new waves appeared, which was due to the formation of a supramolecular complex of ARS with LYS in the solution. The stoichiometry of the ARS-LYS complex was further calculated by the electrochemical data with the results of the binding ratio as 3 : 1 and the binding constant as $2.82 \times 10^{14}$. Under the selected conditions, the decrease of the second order derivative linear sweep voltammetric reductive peak current of ARS was in proportion to the LYS concentration in the range from 0.8 to 35.0 mg/L and the detection limit of LYS was calculated as 0.52 mg/L (3𝜎). Different kinds of LYS samples were detected satisfactorily with this method.

Direct electrochemistry of hemoglobin (Hb) entrapped in the dextran (De) film on the surface of a room temperature ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate (BMIMPF₆) modified carbon paste electrode (CILE) has been investigated. UV-Vis and FT-IR spectroscopy showed that Hb retained its native structure in the De film. Scanning electron microscopy (SEM) indicated an uniform film was formed on the electrode surface. Cyclic voltammetric experiments indicated that the electron transfer efficiency between Hb and the electrode was greatly improved due to the presence of the De film and ionic liquid, which provided a biocompatible and higher conductive interface. A pair of well-defined and quasi-reversible redox peak was obtained with the anodic and cathodic peaks located at -0.195 V and -0.355 V in pH 7.0 phosphate buffer solution, respectively. The electrochemical parameters were calculated by investigating the relationship of the peak potential with the scan rate. The fabricated De/Hb/CILE showed good electrocatalytic ability to the reduction of H₂O₂ with the linear concentration range from $4.0 \times 10^{-6}$ to $1.5 \times 10^{-5}$ mol/L and the apparent Michaelis-Menten constant ($K_M^{\text{app}}$) for the electrocatalytic reaction was calculated as 0.17 𝜇M.