Bioelectrochemical Determination of Citric Acid in Real Samples Using a Fully Automated Flow Injection Manifold

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Abstract
An enzymic method for the determination of citric acid in fruits, juices and sport drinks is proposed. The method is based on the action of the enzymes citrate lyase, oxaloacetate decarboxylase and pyruvate oxidase, which convert citric acid into H2O2 with the latter being monitored amperometrically with a H2O2 probe. The enzymes pyruvate oxidase and oxaloacetate decarboxylase were immobilized. A multi-membrane system, consisting of a cellulose acetate membrane for the elimination of interferants, an enzymic membrane and a protective polycarbonate membrane were placed on a Pt electrode and used with a fully automated flow injection manifold. Several parameters were optimized, resulting in a readily constructed and reproducible biosensor. Interference from various compounds present in real samples was minimized. Calibration graphs were linear over the range 0.01–0.9 mM pyruvate, 0.015–0.6 mM oxaloacetate and 0.015–0.5 mM citrate. The throughput was 30 samples h– 1 with an RSD of 1.0% (n = 8); the mean relative error was 2.4% compared with a standard method. The recovery was 96–104%. A 8–10% loss of the initial activity of the sensor was observed after 100–120 injections.

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