Development of an enzyme-modified carbon paste electrode for determining inhibitors of lipoxygenase

In this study, a 15-lipoxygenase-modified carbon paste electrode (15-LOX-MCPE) was developed in connection with the help of voltammetry, which can be used as an assay system for screening drugs with inhibiting lipoxygenase (LOX) activity. The influence of different experimental conditions (LOX loading of carbon paste, pH, type of buffer system etc.) was investigated in order to optimise the biosensing device. The best composition of the biosensor is 30% paraffin oil, 68% graphite powder and 2% LOX. The optimised voltammetric measurement medium is Sörensen/NaOH (0.1M, pH 9.0) using linoleic acid as a substrate. Under these conditions the hydroperoxy linoleic acid is formed, which can be oxidised at a potential of +0.9 V versus Ag/AgCl/3M KCl. The applicability of the LOX biosensor as assay of lipoxygenase inhibitors was successfully tested with nordihydroguaiaretic acid, zileuton and fenleuton, which are well known inhibitors of LOX.