Related Experiment Video
Updated: May 4, 2026
Quantitative Determination of De Novo Fatty Acid Synthesis in Brown Adipose Tissue Using Deuterium Oxide
Published on: May 12, 2023
Optimized method for quantification of total F(2)-isoprostanes using gas chromatography-tandem mass spectrometry
David R Briskey1, Gary R Wilson2, Robert G Fassett2
1Exercise and Oxidative Stress Research Group, School of Human Movement Studies, University of Queensland, Brisbane, Australia; School of Medicine, University of Queensland, Brisbane, Australia.
Abstract:
F2-isoprostanes are produced from the oxidative degradation of arachidonic acid and are considered the gold standard marker of lipid peroxidation in biological samples. We developed a liquid-liquid extraction method for the determination of total isoprostanes using negative chemical ionization gas chromatography-tandem mass spectrometry in plasma and tissue homogenates. Incorporating liquid-liquid extraction allows for greater sample through-put than current approaches. Here we describe the protocol and include numerous trouble-shooting suggestions. The method found healthy individuals with 150-250 pg of isoprostanes per ml of plasma and end stage kidney disease patients to have the highest measured values of up to 1100 pg/ml. This assay has an accurate working linear range of 40-1000 pg of isoprostanes (100-2500 pg/ml) and an average coefficient of variance of 7%. Tissue values for healthy mice liver were 50-70 pg/μg protein. This method provides increased ion selectivity and detection capabilities with economical sample through-put.
