Abstract
Lipid mediators, small molecules involved in regulating inflammation and its resolution, are a class of lipids of wide interest as their levels in blood and tissues may be used to monitor health and disease states or the effect of new treatments. These molecules are present at low levels in biological samples, and an enrichment step is often needed for their detection. We describe a rapid and selective method that uses new low-cost molecularly imprinted (MIP) and non-imprinted (NIP) polymeric sorbents for the extraction of lipid mediators from plasma and tissue samples. The extraction process was carried out in solid-phase extraction (SPE) cartridges, manually packed with the sorbents. After extraction, lipid mediators were quantified by liquid chromatography–tandem mass spectrometry (LC–MSMS). Various parameters affecting the extraction efficiency were evaluated to achieve optimal recovery and to reduce non-specific interactions. Preliminary tests showed that MIPs, designed using the prostaglandin biosynthetic precursor arachidonic acid, could effectively enrich prostaglandins and structurally related molecules. However, for other lipid mediators, MIP and NIP displayed comparable recoveries. Under optimized conditions, the recoveries of synthetic standards ranged from 62% to 100%. This new extraction method was applied to the determination of the lipid mediators concentration in human plasma and mouse tissues and compared to other methods based on commercially available cartridges. In general, the methods showed comparable performances. In terms of structural specificity, our newly synthesized materials accomplished better retention of prostaglandins (PGs), hydroxydocosahexaenoic acid (HDoHE), HEPE, hydroxyeicosatetraenoic acids (HETE), hydroxyeicosatrienoic acid (HETrE), and polyunsaturated fatty acid (PUFA) compounds, while the commercially available Strata-X showed a higher recovery for dihydroxyeicosatetraenoic acid (diHETrEs). In summary, our results suggest that this new material can be successfully implemented for the extraction of lipid mediators from biological samples.
Highlights
Lipid mediators are low abundant signaling molecules mainly originated from arachidonic acid (ArA), eicosapentaenoic acid (EPA), or docosahexaenoic acid (DHA) under the catalysis of cyclooxygenase (COX), lipoxygenase (LOX), or cytochrome P450 (CYP)
We investigated the ability of the newly synthesized sorbents to enrich a mixture of prostaglandin standards from acidic buffers
This work demonstrates the applicability of new molecularly imprinted (MIP)/NIP-based solid-phase extraction (SPE) for the selective extraction of lipid mediators from human plasma and mouse tissues
Summary
Lipid mediators are low abundant signaling molecules mainly originated from arachidonic acid (ArA), eicosapentaenoic acid (EPA), or docosahexaenoic acid (DHA) under the catalysis of cyclooxygenase (COX), lipoxygenase (LOX), or cytochrome P450 (CYP). According to their precursors, lipid mediators can be divided into two categories, omega-6 (ω-6) and omega-3 (ω-3). A quantitative analysis of lipid mediators in biological samples is the key to a mechanistic understanding of their biological roles and is usually achieved by liquid chromatography–mass spectrometry (LC–MSMS) [6,7]
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