Abstract

Oxylipins derived from omega-3 and -6 fatty acids are actively involved in inflammatory and immune processes and play important roles in human disease. However, as the interest in oxylipins increases, questions remain regarding which molecules are detectable in plasma, the best methods of collecting samples, and if molecules are stable during collection and storage. We thereby built upon existing studies by examining the stability of an expanded panel of 90 oxylipins, including specialized pro-resolving lipid mediators (SPMs), in human plasma (n = 5 subjects) during sample collection, processing, and storage at −80 °C. Oxylipins were quantified using liquid chromatography-tandem mass spectrometry (LC/MS/MS). Blood samples collected in ethylenediaminetetraacetic acid (EDTA) or heparin followed by up to 2 h at room temperature prior to processing showed no significant differences in oxylipin concentrations compared to immediately processed samples, including the SPMs lipoxin A4 and resolvin D1. The majority of molecules, including SPMs, remained stable following storage for up to 1 year. However, in support of previous findings, changes were seen in a small subset of oxylipins including 12-HETE, TXB2, 14-HDHA, and 18-HEPE. Overall, this study showed that accurate measurements of most oxylipins can be obtained from stored EDTA or heparin plasma samples using LC/MS/MS.

Highlights

  • Oxylipins are endogenously generated from omega-3 and -6 polyunsaturated fatty acids (PUFAs) via enzymatic and non-enzymatic oxidation [1]

  • ethylenediaminetetraacetic acid (EDTA) or heparin plasma samples kept at room temperature for two hours showed no statistically significant differences in oxylipin concentrations by delayed processing time (Table 2 for EDTA and Table 3 for heparin)

  • EDTA and heparin samples after 2 h of delayed plasma processing at room temperature showed no significant differences in oxylipin concentrations compared to an immediately processed sample

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Summary

Introduction

Oxylipins are endogenously generated from omega-3 and -6 polyunsaturated fatty acids (PUFAs) via enzymatic and non-enzymatic oxidation [1]. These bioactive metabolites are the mediators of PUFA effects on inflammatory and immune processes, including the resolution of inflammation and reduction in pain [2]. Recent developments in targeted lipidomic methodologies have made the identification and quantification of oxylipins more reliable [3,4,5,6]. Quantifying oxylipins using lipidomics may lead to a deeper understanding of the contribution of these bioactive metabolites in human disease. Longitudinal epidemiologic cohort studies with repositories of bio-specimens offer a potentially important opportunity to investigate the role of oxylipins in the development and progression of human diseases

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