A Protocol for Quantifying Lipid Peroxidation in Cellular Systems by F2-Isoprostane Analysis
Figure 2
F2-isoprostanes are formed from peroxidation of arachidonic acid.
(A-C) Copper-induced in vitro peroxidation of arachidonic acid resulted in the formation of three F2-isoprostane isomers. Arachidonyl PAF-C16 was subjected to 50 µM of CuCl2, with or without BHT, for the indicated time points and the levels of 8-isoPGF2α-III; 5-iPF2α-VI and 8,12-iso-iPF2α-VI were measured by UPLC-MS/MS. (D-F) F2-isoprostanes were formed in a time-dependent manner in cell lysates upon copper induced lipid peroxidation. HepG2 cell lysates were incubated with CuCl2 for indicated time points in the presence or absence of Trolox or BHT. Levels of 8-isoPGF2α-III; 5-iPF2α-VI and 8,12-iso-iPF2α-VI were analyzed in time by UPLC-MS/MS. Data are represented as mean ± S.D. (G) UPLC-MS/MS chromatogram overlay of extracted F2-isoprostanes of cells treated with GOX for indicated times.