DETERMINATION OF ALDEHYDIC LIPID-PEROXIDATION PRODUCTS WITH DABSYLHYDRAZINE BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A high-performance liquid chromatographic method has been developed to measure adlehyde products of lipid peroxidation including n-alkanals, trans-2-alkenals, 4-hydroxy-2-nonenal, and malonaldehyde. The chromophoric reagent dabsylhydrazine reacts with various aldehydes produced from two different lipid peroxidation model systems: Fe2+/H2O2 and Fe3+/vitamin C oxidation of arachidonic and linoleic acids, respectively. The formed products were carefully characterized by IR, MS, and NMR spectral analysis and proved to be hydrazone rather than 2-pyrazoline or pyrrole derivatives. EI-MS of all the derivatized straight-chain aldehydes showed a characteristic (M - 28) fragment in the spectra with high consistency. Physicochemical properties including melting points are also described. The detection limits in the range of 11 pmol (5 ng) have been reached by using an NP Octadecyl C-18 reversed-phase column. 4-Hydroxy-2-nonenal and hexanal were found to be the most abundant aldehydes in the lipid peroxidation systems described. The dose-response curve showed great linearity and exhibited good reproducibility of this procedure for quantitative estimation.