The formation of non-enzymatic oxylipins is catalysed by reactive oxygen species. Reactive oxygen species are produced in response to pathogen attack. In this study, the accumulation of non-enzymatically formed hydroxy fatty acids and F₁-phytoprostanes in leaves of Arabidopsis thaliana (L.) Heyhn upon infection with Pseudomonas syringae was investigated and compared with the accumulation of the enzymatically formed oxylipins jasmonic acid and 12-oxo-phytodienoic acid. Levels of all oxylipins increased after infection with a virulent and with an avirulent strain of P. syringae. Inoculation of the avirulent strain resulted in a biphasic accumulation with a first maximum around 5 h which was missing after inoculation of the virulent strain. Levels of free and esterified hydroxy fatty acids and F₁-phytoprostanes increased after pathogen treatment; however, esterified compounds were 30 times more abundant than free oxylipins. The increase of the free compounds occurred later than the increase of the esterified compounds suggesting that non-enzymatic lipid oxidation occurs predominantly in membranes from which oxidised lipids can be released.