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 labs assessed the technical variability and comparability of 133 oxylipins using a harmonized and standardized protocol, common biological materials (i.e. 7 quality control plasmas), standard calibration series and analytical methods. The quantitative analysis is based on a standard calibration series with isotopically labelled 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 labs was examined using consensus value estimates and unsupervised /supervised multivariate analysis (i.e. PCA and PLS-DA). Inter-lab 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 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.

中文翻译：

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统一的程序导致实验室间总oxylipins 的可比量化。

Oxylipins 是有效的脂质介质，参与多种生理过程。他们的分析有可能提供有关人类健康和疾病的大量信息，并且是转化为临床应用的有前途的技术。然而，独立组产生的结果很少具有可比性，这增加了实施国际商定协议的必要性。我们对基于 MS 的总 oxylipins 定量分析进行了实验室间比较。五个独立实验室使用统一和标准化的方案、常见生物材料（即 7 种质量控制血浆）、标准校准系列和分析方法评估了 133 种 oxylipins 的技术可变性和可比性。定量分析基于具有同位素标记的内标的标准校准系列。使用标准化方案，73% 的 oxylipins 的技术方差在 ±15% 以内，但是，大多数环氧脂肪酸被确定为关键分析物，因为浓度。使用一致值估计和无监督/监督多变量分析（即 PCA 和 PLS-DA）检查实验室确定的浓度的可比性。实验室间的差异是有限的，不会干扰我们区分不同血浆的能力。此外，所有实验室都能够识别血浆之间的相似差异。总而言之，我们表明，使用标准化的样品制备方案，可以实现较低的技术变异性。