In this work, an electroenzymatic methodology based on two coupled enzymatic activities (glycerokinase and glycerol-3-phosphate oxidase) was developed using an oxygen Clark-type electrode for the determination of free and total glycerol in biodiesel samples. The enzymatic conversion of glycerol consumes oxygen, which is measured amperometrically in a Clark-type electrode and correlated with the concentration of glycerol in the sample. The electroenzymatic method proposed showed a good linear correlation coefficient (R=0.9990) with a linear response in the concentration range of 6.25×10−5 to 6.25×10−4% (w/v) and limits of detection and quantification at 1.0×10−5% and 3.0×10−5% (w/v), respectively. Good correlations were found between the results obtained in this work and those by the gas chromatography technique (R=0.9994). The proposed method was shown to be promising for the analysis of glycerol in biodiesel samples, with a simple and inexpensive methodology compared with the gas chromatography technique.
Electroanalysis 03/2010; 22(9):995 - 999. DOI:10.1002/elan.200900428 · 2.50 Impact Factor