Flow Injection Amperometric System Coupled with a Well-Plate for Fast Screening of Total Antioxidant Capacity

Abstract

ABSTRACTA flow injection (FI) system with an amperometric detection in combination with a 96-well-plate has been developed for high throughput screening of total antioxidant capacity. It is based on the reaction between dichromate and antioxidants Chromium reducing antioxidant capacity (CHROMAC) assay. Antioxidant reduces dichromate leading to a decrease of dichromate concentration, in which reacts further with triiodide reagent. The slow reaction between antioxidant and dichromate takes place in parallel in a 96-well microplate, with the reaction time sufficiently long to reach steady state conditions. Microliter volumes of the remaining dichromate solution were manually injected into the FI amperometric system. Reaction with iodide in acidic medium produces triiodide which undergoes electrochemical reduction on a glassy carbon working electrode at 200 mV versus a Ag/AgCl reference electrode. The decrease of electrical current was directly proportional to the antioxidant capacity. The optimum conditions have been achieved as follow: 160 mg L−1 of potassium dichromate, pH of dichromate solution of 2.82, 0.005 mol L−1 of potassium iodide, 0.20 mol L−1 of hydrochloric acid, and 3.0 mL min−1 total flow rate. The developed method was applied for the determination of total antioxidant capacity of tea and herbal infusions and fruit juice samples. The results obtained indicated that the developed method had good correlation with the batch CHROMAC spectrophotometric method and 1,1-diphenyl-2-picrylhydrazyl scavenging assay. The proposed method provides fast and high throughput analysis, convenient operation, and low consumption of chemical reagents and samples.