Determination of ω-6 and ω-3 PUFA metabolites in human urine samples using UPLC/MS/MS

Abstract

The ω-6 and ω-3 polyunsaturated fatty acids (PUFAs) such as arachidonic acid (AA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) are the precursors of various bioactive lipid mediators including prostaglandins, thromboxanes, leukotrienes, hydroxyeicosatetraenoic acid, isoprostanes, lipoxins, and resolvins (Rvs). These lipid mediators play important roles in various physiological and pathological processes. The quantitative determination of PUFA metabolites seems necessary for disease research and for developing biomarkers. However, there is a paucity of analytical methods for the quantification of ω-6 and ω-3 PUFA metabolites—the specialized pro-resolving mediators (SPMs) present in the human urine. We developed a method for the quantification of ω-6 and ω-3 PUFA metabolites present in human urine using ultra-performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS). The developed method shows good linearity, with a correlation coefficient >0.99 for all of the analytes. The validation results indicate that our method is adequately reliable, accurate, and precise. The method was successfully used to examine urine samples obtained from 43 healthy volunteers. We could identify 20 PUFA metabolites, and this is the first report of the quantitative determination of RvD1, 17(R)-RvD1, 11-dehydro thromboxane B3, RvE2, and 5(S)-HETE in human urine. The urinary 8-iso PGF(2α) and PGE2 levels were significantly higher in the men smokers than in the men nonsmokers (p < 0.05). In this study, we developed an accurate, precise, and novel analytical method for estimating the ω-6 and ω-3 PUFA metabolites, and this is the first report that the SPMs derived from EPA and DHA are present in human urine.

Lipoxin A4 Attenuates Obesity-Induced Adipose Inflammation and Associated Liver and Kidney Disease

“…Previous work demonstrates that LXA 4 increases adiponectin in adipose explants (Claria et al, 2012), is present in human adipose tissue (Claria et al, 2013) and urine (Sasaki et al, 2015) and herein we observed a trend towards increased WAT adiponectin production in LX-treated mice (Figure 1D). Therefore, we hypothesized that LXs may mediate protection in our obesity model via induction of adiponectin in WAT.…”

“…LXA 4 potently attenuate acute inflammation (Maderna and Godson, 2009) and age-associated adipose inflammation ex vivo (Borgeson et al, 2012). Of note SPM have been identified in a number of human tissues and fluids including spleen, lymph nodes (Colas et al, 2014), urine (Sasaki et al, 2015) and WAT (Claria et al, 2013). Importantly, whether LXA 4 actively attenuates chronic inflammation remains to be addressed.…”

Cross-Classification of Human Urinary Lipidome by Sex, Age, and Body Mass Index

“…Of the 46 metabolites scanned in this target lipidomic analysis (S2 Table), four PUFAs and 25 oxylipins were observed at detectable and quantifiable levels in human urine. Similar to other studies [21], 9-hydroxyoctadecadienoic acid (HODE), 9-oxooctadecadienoic acid (KODE), 12- hydroxyeicosatetraenoic acid (HETE), docosahexaenoic acid (DHA), arachidonic acid (AA), and prostaglandin (PG) derivatives were detected at relatively high levels among the monitored ions (S6 Table). …”

Bioactive products formed in humans from fish oils

“…There are few data attesting to their formation in humans based on rigorous mass spectrometric methodology. Most clinical investigations reporting the formation of SPMs rely on immunoassays ( 21 ) or MS without synthetic internal standards structurally similar to the SPMs under investigation (22)(23)(24)(25)(26)(27)(29)(30)(31)(32), often in ex vivo stimulated systems ( 17,(33)(34)(35)(36).…”