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Assay of liver cytosol lipoxygenase by differential pulse polarography
1Department of Biochemistry, Faculty of Medicine, National University of Singapore, Republic of Singapore.
Abstract:
Lipoxygenase activity assayed by differential pulse polarography was found to be more sensitive than that determined by the increase in absorption at 234 nm. The lipoxygenase activity level in the liver cytosol of rats fed oxidized palm oil was significantly higher than that of the control animals fed either saline or fresh palm oil. The effects of flavonoids on the inhibition of lipoxygenase activity level in liver cytosol of rats were in the decreasing order quercetin greater than myricetin greater than morin greater than phloretin. The observed free malonaldehyde (MDA) in liver cytosol of rats determined by high-performance liquid chromatography using malonaldehyde-dinitro-phenylhydrazine complex was 23, 25, and 51.2% for rats fed saline, fresh palm oil, and oxidized palm oil, respectively. A linear relationship between the lipoxygenase activity and the free liver cytosol MDA was shown. The assay of lipoxygenase by differential pulse polarography provides a simple, sensitive, and quantitative method for the study of liver lipid peroxidation.