The electrooxidation behavior of 3, 3′,5, 5′-tetramethylbenzidine(TMB) was investigated using a platinum minigrid optically transparent thin-layer spectroelectrochemical cell. TMB underwent one two-electron electrooxidation process to yield quinonediimine in the pH range from 2.0 to < 4.0, and two consecutive one-electron electrooxidation processes, gave the mediate product free radical of TMB first, then gave the oxidation product quinonediimine in the pH range from 4.0 to < 7.0. In the pH range from 7.0 to 10.0, the electrooxidation of TMB was also one two-electron electrooxidation process to yield an azo compound. The formal potential E0'and the electron transfer number of the electrooxidation of TMB at pH 2.0 and pH 8.4 were determined by spectroelectrochemical techniques.
A new quantitative determination method of proteins using beryllon ?by voltammetric technique was developed in this paper. In pH 3.5 Britton-Robinson (B-R) buffer solution, beryllon ?can bind with human serum albumin (HSA) to form an electro-inactive supermolecular complex. Beryllon ?has a well-defined voltammetric reduction peak at 0.38 V (vs. SCE) and the addition of protein in this solution can cause the decrease of the reductive peak current. Based on the decrease of the reduction peak current, a new electrochemical method for the determination of HSA was established with linear range of 1.0~40.0 mg/L and the detection limit of 1.0 mg/L. This method is further applied to the determination of real sample of healthy human serum.
