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Harmonized procedures lead to comparable quantification of total oxylipins across laboratories.

Abstract：

: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 133 oxylipins using a harmonized and standardized protocol, common biological materials (i.e., seven quality control plasmas), standard calibration series, and analytical methods. The quantitative analysis was based on a standard calibration series with isotopically labeled internal standards. Using the standardized protocol, the technical variance was within ±15% for 73% of oxylipins; however, most epoxy fatty acids were identified as critical analytes due to high variabilities in concentrations. The comparability of concentrations determined by the laboratories was examined using consensus value estimates and unsupervised/supervised multivariate analysis (i.e., principal component analysis and partial least squares discriminant analysis). 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.

journal_name

J Lipid Res

journal_title

Journal of lipid research

authors

Mainka M,Dalle C,Pétéra M,Dalloux-Chioccioli J,Kampschulte N,Ostermann AI,Rothe M,Bertrand-Michel J,Newman JW,Gladine C,Schebb NH

doi

10.1194/jlr.RA120000991

subject

Has Abstract

pub_date

2020-11-01 00:00:00

pages

1424-1436

issue

eissn

0022-2275

issn

1539-7262

pii

jlr.RA120000991

journal_volume

pub_type

杂志文章

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Harmonized procedures lead to comparable quantification of total oxylipins across laboratories.