Abstract
A L-ascorbic acid biosensor based on ascorbate oxidase has been developed. The enzyme wasextracted from the mesocarp of cucumber (Cucumis sativus) by using 0.05 mol L -1 phosphate buffer, pH5.8 containing 0.5 mol L -1 NaCl. After the dialysis versus phosphate buffer0.05 mol L -1 pH 5.8, the enzyme was immobilized onto nylon net through glutaraldehyde covalentbond. The membrane was coupled to an O 2 electrode and the yielding reaction monitored by oxygendepletion at -600 mV using flow injection analysis optimized to 0.1 mol L -1 phosphate buffer pH 5.8, as thecarrier solution and flow-rate of 0.5 mL min -1 . The ascorbic acid calibration curve was linear from 1.2x10 -4 to1.0x10 -3 mol L -1 . The evaluation of biosensor lifetime leads to 500 injections. Commercial pharmaceuticalsamples were analyzed with the proposed method and the results were compared with those obtained byhigh-performance liquid chromatography (HPLC).
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