Determination of Malic Acid in Real Samples by Using Enzyme Immobilized Reactors and Amperometric Detection

Abstract

The malate dehydrogenase (MDH) and ascorbate oxidase were immobilized independently, onto silanized controlled porous silica and packed in a tygon tube. The reactors were inserted in the flow system, and the malic acid was determined by measurement of NADH produced by enzymatic reaction. The NADH was reoxidized in a wall jet cell that consisted of spectrographic graphite, Ag/AgCl, KCl_(sat), and steel needle as work, reference, and counter electrodes, respectively. The current intensities were measured at 390 mV. The malate calibration curve shows a linear range from 5.0 × 10⁻⁶ to 1.0 × 10⁻⁴ mol L⁻¹, the lifetime was 40 analyses, after that a decrease of 20% on the response is observed. Three different citric juices were analyzed and a good correlation between the proposed method and spectrophotometric commercial kit were obtained.

Keywords:

• Citrus juice
• Malic acid
• Malate dehydrogenase
• Ascorbic acid

Acknowledgment

The authors thank FAPESP and EU for the financial support and A. Manzoli thanks CNPq for the fellowship.