Peroxidative stimuli mediated by high polyunsaturated fatty acid administration in rats induced in vivo lipid peroxidation and resulted in increased urinary excretion of a number of lipophilic aldehydes and related carbonyl compounds. These secondary lipid peroxiation products, measured as 2, 4-dinitrophenylhydrazine deritives, were detected and identified by the newly developed HPLC method. The identified urinary lipophilic nonpolar aldehydes and related carbonyl compounds were butanal, butan-2-one, pentan-2-one, hexanal, hex-2-enal, hepta-2, 4-dienal, hept-2-enal, octanal, and oct-2-enal. Lipophilic polar aldehydes such as 4-hydroxyhex-2-enal and 4-hydroxyoct-2-enal were also identified. A polyunsaturated fatty acid diet containing n-3 fatty acids generally caused high levels of urinary excretion of lipophilic aldehydes and related carbonyl compounds in rats than a normal diet. Significantly increased secondary lipid peroxidation products were hexanal, hepta-2, 4-dienal, octanal, 4-hydroxyhex-2-exal, 4-hydroxyoct-2-enal, and a number of unidentified compunds.