Abstract
A specific and sensitive high performance liquid chromatographic method using immobilized enzyme membrane electrode is described for the simultaneous determination of ethanol and glucose.
Photo cross-linked polymer (PVA-SbQ) membrane coimmobilized alcohol oxidase and glucose oxidase was attached on the platinum electrode in the flow cell. The flow cell with the immobilized enzyme membrane was positioned after the column in HPLC. Phosphate buffer solution (pH 7.0, 0.05 mol/dm3)was used as a mobile phase and was flowed at a rate of 0.8 ml/min. At first, glucose was eluted through the HPLC column at ca.12 min and enzymically oxidized to gluconolactone and hydrogen peroxide by glucose oxidase in the immobilized enzyme membrane. Next, ethanol was eluted from HPLC column at ca.14 m in and oxidized to acetaldehyde and hydrogen peroxide by alcohol oxidase at the same enzym e membrane. Finally, produced hydrogen peroxide was amperometrically detected by the electrochemical method. The minimum detectable amounts of the ethanol and glucose were found to be 30 ng and 50 ng, respectively. The combination of specific immobilized enzyme membrane and electrochemical method enabled to perform highly selective detection of ethanol and glucose. This method was successfully applied for the determination of ethanol and glucose in alcoholic beverages.
