期刊
ANALYTICAL CHEMISTRY
卷 87, 期 23, 页码 11721-11731出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.5b02794
关键词
-
资金
- Technologie Servier
- Technologie Servier through the Imperial College Stratified Medicine Graduate Training Program in Systems Medicine and Spectroscopic Profiling (STRATi-GRAD)
- UK Medical Research Council (MRC) [MC_PC_12025]
- Bruker Biospin
- Waters Corporation
- Metabometrix
- Imperial College
- National Institute for Health Research (NIHR) (England)
- Medical Research Council [MC_PC_12025] Funding Source: researchfish
- MRC [MC_PC_12025] Funding Source: UKRI
Lipid mediators, highly bioactive compounds synthesized from polyunsaturated fatty acids (PUFAs), have a fundamental role in the initiation and signaling of the inflammatory response. Although extensively studied in isolation, only a limited number of analytical methods offer a comprehensive coverage of the oxylipin synthetic cascade applicable to a wide range of human biofluids. We report the development of an ultrahigh-performance liquid chromatography-electrospray ionization triple quadrupole mass spectrometry (UHPLC-MS) assay to quantify oxylipins and their PUFA precursors in 100 mu L of human serum, plasma, urine, and cell culture supernatant. A single 15 min UHPLC run enables the quantification of 43 oxylipins and 5 PUFAs, covering pro and anti-inflammatory lipid mediators synthesized across the cyclooxygenase (COX), lipoxygenase (LOX), and cytochrome P450 (CYP450) pathways. The method was validated in multiple biofluid matrixes (serum, plasma, urine, and cell supernatant) and suppliers, ensuring its suitability for large scale metabonomic studies. The approach is accurate, precise, and reproducible (RSD < 15%) over multiple days and concentrations. Very high sensitivity is achieved with limits of quantification inferior to picograms for the majority of analytes (0.05-125 pg) and linear range spanning up to 5 orders of magnitude. This enabled the quantification of the great majority of these analytes at their low endogenous level in human biofluids. We successfully applied the procedure to individuals undergoing a fasting intervention; oxylipin profiles highlighted significantly altered PUFA and inflammatory profiles in accordance with previously published studies as well as offered new insight on the modulation of the biosynthetic cascade responsible for the regulation of oxylipins.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据