Official websites use. Share sensitive information only on official, secure websites. Oxylipins are potent lipid mediators involved in a variety of physiological processes. Their profiling has the potential to provide a wealth of information regarding human health and disease and is a promising technology for translation into clinical applications. However, results generated by independent groups are rarely comparable, which increases the need for the implementation of internationally agreed upon protocols.
We performed an interlaboratory comparison for the MS-based quantitative analysis of total oxylipins. Five independent laboratories assessed the technical variability and comparability of oxylipins using a harmonized and standardized protocol, common biological materials i.
The quantitative analysis was based on a standard calibration series with isotopically labeled internal standards. Interlaboratory variability was limited and did not interfere with our ability to distinguish the different plasmas. Moreover, all laboratories were able to identify similar differences between plasmas.
In summary, we show that by using a standardized protocol for sample preparation, low technical variability can be achieved. Harmonization of all oxylipin extraction and analysis steps led to reliable, reproducible, and comparable oxylipin concentrations in independent laboratories, allowing the generation of biologically meaningful oxylipin patterns. Keywords: eicosanoids, lipidomics, liquid chromatography, mass spectrometry, quantitation, harmonization, plasma lipids, interlaboratory comparison, oxidized fatty acids, lipid mediators.
Eicosanoids and other oxylipins are potent lipid mediators produced via the oxygenation of PUFAs. PUFAs can be oxygenated enzymatically by cyclooxygenases to form prostanoids, by lipoxygenases to form hydroperoxy fatty acids, which react further to mono- and poly-hydroxylated fatty acids, or by cytochrome P monooxygenases giving rise to epoxy and hydroxy fatty acids, or nonenzymatically by free radicals during autoxidation 1 , 2.
