Comparison of quantitative eicosanoids profiling in human plasma and serum by ultra-fast multiple reaction monitoring

Significance of the Topic

Eicosanoid profiling by multiple reaction monitoring is essential for understanding physiological functions and identifying disease biomarkers. Comparing plasma and serum profiles reveals how sample preparation and anticoagulants influence measured metabolic patterns.

Objectives and Study Overview

This study aimed to quantitatively compare 68 detected eicosanoids and related fatty acid metabolites across EDTA plasma, heparin plasma, and serum. Out of an initial panel of 196 targets, the method consistently quantified metabolites in a 20-minute reversed-phase LC-MS run.

Methodology and Instrumentation

Human plasma and serum samples were subjected to protein precipitation with formic acid/methanol, followed by solid-phase extraction. Analytes were analyzed in triplicate using fast polarity switching for selected species.

• Extraction: STRATA-X 10 mg SPE cartridge
• Chromatography: Kinetex C8 column (2.1 × 150 mm, 2.6 μm)
• Mass spectrometry: Nexera UHPLC coupled to LCMS-8060NX triple quadrupole
• Monitoring: 190 standards plus 18 isotope-labeled internal standards

Main Results and Discussion

EDTA and heparin plasma samples yielded detection of 67 eicosanoids, whereas serum samples contained 44. Thromboxane B2 was absent in plasma due to anticoagulant suppression, while cyclooxygenase products (e.g., PGE2, PGD2) and lipoxygenase metabolites (e.g., HETEs, leukotrienes) were elevated in plasma compared to serum. Cytochrome P450-derived dihydroxy species showed minimal variation among matrices.

Benefits and Practical Applications

This streamlined MRM-LC-MS workflow delivers rapid, reproducible quantification of key eicosanoids, supporting clinical research, quality control, and biomarker discovery through high-throughput metabolic profiling.

Future Trends and Potential Applications

Future directions include expanding target panels, integrating automated data analysis with metabolic maps for real-time interpretation, and applying this approach to patient cohorts for personalized diagnostics. Advances in high-resolution MS and ion mobility may further resolve isomeric species.

Conclusion

The developed ultra-fast MRM method enabled comprehensive detection of 68 eicosanoids in human blood matrices and visualization of quantitative differences via metabolic mapping, highlighting sample-specific enzymatic activities.

Reference

Yamada M. et al. J. Chromatography B, 995, 74-84 (2015).