Oxylipins, a subclass of lipid mediators, are metabolites of various polyunsaturated fatty acids with crucial functions in regulation of systemic inflammation. Elucidation of their roles in pathological conditions requires accurate quantification of their levels in biological samples. We refined an ultra-performance liquid chromatography-multiple reaction monitoring-mass spectrometry (UPLC-MRM-MS)-based workflow for comprehensive and specific quantification of 131 endogenous oxylipins in human plasma, in which we optimized LC mobile phase additives, column, and gradient conditions. We employed heatmap-assisted strategy to identify unique transitions to improve the assay selectivity and optimized solid phase extraction procedures to achieve better analyte recovery. The method was validated according to FDA guidelines. Overall, 94.4% and 95.7% of analytes at tested concentrations were within acceptable accuracy (80-120%) and precision (CV < 15%), respectively. Good linearity for most analytes was obtained with R2 > 0.99. The method was also validated using a standard reference material-SRM 1950 frozen human plasma to demonstrate inter-lab compatibility. Graphical abstract ᅟ.

中文翻译：

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使用反相液相色谱-三重四极杆质谱联用热图辅助选择跃迁对人血浆中的磷脂进行全面分析。

脂蛋白是脂质介体的一个亚类，是多种多不饱和脂肪酸的代谢产物，在调节全身性炎症中起关键作用。阐明其在病理条件下的作用需要准确定量其在生物样品中的水平。我们优化了基于超高效液相色谱-多反应监测-质谱（UPLC-MRM-MS）的工作流程，以对人血浆中的131种内源性脂蛋白进行全面而特定的定量，其中我们优化了LC流动相添加剂，色谱柱和梯度条件。我们采用了热图辅助策略来识别独特的跃迁，以提高测定的选择性，并优化了固相萃取程序，以实现更好的分析物回收率。该方法已根据FDA指南进行了验证。总体而言，分别为94.4％和95。在测试浓度下，有7％的分析物分别在可接受的准确度（80-120％）和精密度（CV <15％）之内。对于R2> 0.99，大多数分析物具有良好的线性。该方法还使用标准参考材料-SRM 1950冷冻人血浆进行了验证，以证明实验室间的兼容性。图形摘要ᅟ。