Oxylipin Quantification and Pathway Profiling

Oxylipins are key regulators of inflammation, stress response, and immune signaling. At Creative Proteomics, we provide targeted and custom oxylipin profiling using high-resolution LC–MS/MS to distinguish isomeric species and ensure accurate quantification.

Our validated workflows help researchers uncover lipid mediator dynamics across biofluids, tissues, and plant systems—delivering reproducible, publication-ready results.

We Solve:

  • Poor differentiation of isomeric oxylipins
  • Instability of oxygenated fatty acids during handling
  • Incomplete quantification across complex matrices

Our Advantages:

  • UHPLC–MS/MS and HRAM PRM verification
  • Matrix-specific extraction preserving redox integrity
  • >50 isotope-labeled internal standards for accuracy
Request Analysis

Submit Your Request Now

×
Boston University
University at Buffalo
UMass Medical School
Hawaii Pacific University
Medizinische Fakultät
Louisiana State University
Nyulangone
Nature's Fynd
Dietary Supplement Manufacturer
  • Solution We Provide
  • Detectable Species
  • Advantages
  • Workflow
  • Methods
  • Results and Data Analysis
  • Sample Requirements
  • FAQ

What Are Oxylipins?

Oxylipins are bioactive lipids formed by oxygenation of polyunsaturated fatty acids . They include eicosanoids, specialized pro-resolving mediators, isoprostanes, and plant jasmonates. These molecules regulate inflammation, vascular tone, pain, immune signaling, and stress responses. Because many species are isomeric and unstable, accurate quantification requires rigorous sample handling and targeted mass spectrometry.

Oxylipin Profiling Services at Creative Proteomics

Targeted Quantification Studies

Use predefined oxylipin panels to quantify key lipid mediators—ideal for cross-study comparisons or biomarker verification.

Custom Pathway Panels

Tailor your assay to specific biosynthetic branches (COX, LOX, CYP) or signaling mediators relevant to your model.

Treatment-Response Profiling

Quantify oxylipin shifts across different experimental conditions, treatments, or exposures to map biological response.

Time-Course & Kinetic Tracking

Capture the dynamics of lipid mediator release with sampling strategies designed for temporal resolution.

Cross-Matrix Translation Projects

Analyze matched samples across biofluids, tissues, and cells—human, animal, or plant—to assess systemic signaling.

Plant Oxylipin Stress Packages

Quantify jasmonates and related plant lipid signals under abiotic or biotic stress, optimized for diverse crop matrices.

Full List of Detectable Oxylipins in Our LC–MS/MS Assay

The panel is customizable by matrix and study goals. Below is a representative, validated-or-commonly-validated list for oxylipin analysis by LC–MS(/MS). Availability may vary by matrix and standards.

Family / PathwayRepresentative Analytes (non-exhaustive but practical)
Prostaglandins (COX)PGE2, PGD2, PGF2α, PGF3α, 6-keto-PGF1α (PGI2 metabolite), 13,14-dihydro-15-keto-PGE2, tetranor-PGEM
Thromboxanes (COX)TXB2, 2,3-dinor-TXB2
Leukotrienes (5-LOX)LTB4, 20-OH-LTB4, 20-COOH-LTB4, LTC4, LTD4, LTE4
Lipoxins / SPMsLXA4, LXB4
Resolvins (SPMs)RvE1, RvE2, RvE3; RvD1, RvD2, RvD3, RvD5
Protectins & Maresins (SPMs)PD1/NPD1, PDX; MaR1
EETs (CYP epoxygenase)5,6-EET, 8,9-EET, 11,12-EET, 14,15-EET
DHETs (sEH products)5,6-DHET, 8,9-DHET, 11,12-DHET, 14,15-DHET
HETEs (LOX/CYP)5-HETE, 8-HETE, 9-HETE, 11-HETE, 12-HETE, 15-HETE, 16-HETE, 18-HETE, 19-HETE, 20-HETE
Oxidized AA (oxo-ETEs)5-oxo-ETE, 12-oxo-ETE, 15-oxo-ETE
EPA-derived HEPEs5-HEPE, 8-HEPE, 9-HEPE, 12-HEPE, 15-HEPE, 18-HEPE
DHA-derived HDoHEs/HDHAs4-, 7-, 10-, 11-, 13-, 14-, 17-HDoHE (17-HDHA)
Linoleate oxylipins9-HODE, 13-HODE, 9-oxoODE, 13-oxoODE, 9,10-EpOME, 12,13-EpOME, 9,10-DiHOME, 12,13-DiHOME
α-Linolenate oxylipins9-HOTrE, 13-HOTrE, 9-oxoOTrE, 13-oxoOTrE
Isoprostanes (non-enzymatic)8-iso-PGF2α, 8-iso-PGE2, 2,3-dinor-8-iso-PGF2α, neuroprostanes, isofurans
Nitro-fatty acidsNO2-OA (nitro-oleic acid), NO2-LA (nitro-linoleic acid)
Hydroperoxides (precursors)9-HpODE, 13-HpODE, 5-HpETE, 12-HpETE, 15-HpETE
Other AA metabolites11-, 12-, 15-H-pGF2α (isomeric PGF2α analogs), 15-HETrE
Plant oxylipins (JA pathway)Jasmonic acid (JA), JA-Ile, OPDA (12-oxo-phytodienoic acid), dinor-OPDA, phytoprostanes
Conjugated forms (select)PGE2-glyceryl ester, 13-HODE-G, limited glucuronides/sulfates (upon request)

Why Choose Our Oxylipins Assay?

  • Isomer-Resolved Quantification
    Regio- and stereoisomers (e.g., 5-, 12-, 15-HETE) are separated using high-efficiency UHPLC gradients, avoiding co-elution and misassignment.
  • Stable-Isotope Dilution Accuracy
    Over 50 isotope-labeled internal standards are applied where available, correcting for extraction loss and ion suppression at the compound level.
  • Redox-Safe Sample Handling
    From collection to extraction, samples are protected using antioxidants, chelators, low-light protocols, and cold-chain workflows to limit oxidative artifacts.
  • Cross-Matrix Optimization
    Protocols are tailored for biofluids, cells, tissues, and plants, with validated recovery ranges for each matrix type.
  • Reproducibility Metrics
    Typical intra-assay CVs for quality control pools are ≤15%. Calibration models achieve R2 ≥ 0.995 under standard conditions.
  • Optional High-Resolution Verification
    For identity confirmation or publication support, Orbitrap PRM overlays and ion-ratio consistency can be added to selected targets.

Step-by-Step Workflow for Oxylipin Quantification by LC–MS/MS

Workflow for Oxylipins Analysis

What Methods are Used for Our Oxylipins Analysis?

Chromatography

  • UHPLC: Waters ACQUITY I-Class.
  • Column: C18, 2.1 × 100 mm, 1.7 μm, maintained at 40 °C.
  • Mobile phases: aqueous buffer with low-ionic-strength additive; organic phase acetonitrile/methanol; negative ESI-compatible modifiers.
  • Gradient: sub-15-minute methods for targeted panels; extended gradients for broad coverage.

GC-MS option for select classes

  • GC-MS: Agilent 7890B/5977B, negative chemical ionization.
  • Derivatization: pentafluorobenzyl (PFB) esters for enhanced sensitivity on isoprostanes and related markers.

Mass spectrometry (primary quant)

  • Triple quadrupole: AB Sciex QTRAP 6500.
  • Ionization: ESI-negative; source tuned for thermolabile oxylipins.
  • Acquisition: scheduled MRM with optimized collision energies and dwell times; unit mass resolution.

High-resolution confirmation (as needed)

  • Orbitrap HRAM: Thermo Scientific Orbitrap Exploris 480.
  • Mode: PRM/full scan with sub-ppm mass accuracy; isotopologue checks.
Waters ACQUITY UPLC System

Waters ACQUITY UPLC System (Figure from Waters)

SCIEX Triple Quad™ 6500+

SCIEX Triple Quad™ 6500+ (Figure from Sciex)

Thermo Orbitrap Exploris 480

Orbitrap Exploris 480 (Figure from Thermo)

7890B Gas Chromatograph + 5977 Single Quadrupole

Agilent 7890B-5977B (Figure from Agilent)

Oxylipins Analysis Service: Results and Data Analysis

Standard Deliverables

  • Quantitative Oxylipin Tables — Concentrations for targeted species (e.g., HETEs, EETs, DiHETrEs, EpOMEs, prostaglandins, leukotrienes, resolvins) reported as pg/mL or normalized to input.
  • Pathway-Level Summary — COX, LOX, and CYP450 groupings with ratio readouts to aid interpretation.
  • Normalized Data Matrix — Internal-standard corrected and batch-normalized values for statistics.
  • Raw LC–MS/MS Files — Vendor .raw and open .mzML for full traceability.
  • QC Overview — Recovery, linearity (R2 ≥ 0.995), LLOQ flags, and inter-run RSD% for method confidence.
  • Annotated Chromatograms — EICs and MS/MS spectra supporting compound identity.
Chromatograms, ion-ratio table, and MS/MS spectra confirming isomeric separation and identity of HETEs.

Isomer-Resolved Evidence Panel

Bar plots illustrating oxylipin abundances grouped by biosynthetic pathways: COX, LOX, and CYP450.

Pathway-Level Biological Readout

Advanced Data Analysis (Optional)

  • Multivariate Statistics — PCA/PLS-DA to reveal pattern shifts across conditions.
  • Differential Profiling — Fold-change with FDR-adjusted significance.
  • Enzyme/Pathway Inference — Ratio metrics (e.g., LOX/COX, 5-HETE/15-HETE).
  • Correlation Insights — Associations with phenotypic or biochemical endpoints.

Data Formats

  • Excel/CSV — Clean tables ready for R, MetaboAnalyst, or GraphPad.
  • PDF Result Brief — Distribution plots and QC indicators.
  • Method Appendix — Key analytical settings and internal-standard list for documentation.

Explore our Lipidomics Solutions brochure to learn more about our comprehensive lipidomics analysis platform.

Download Brochure

Sample Requirements for Oxylipins Analysis Solutions

MatrixMinimum amountContainer & preservativeHandling & storageShippingNotes
Plasma/Serum200–500 µLPre-chilled polypropylene; EDTA; optional BHTCollect on ice, quick spin, aliquot, freeze at −80 °CDry iceAvoid hemolysis and repeated freeze–thaw
Urine1–2 mLPolypropylene; optional BHTClarify, aliquot, freeze at −80 °CDry iceNormalize to creatinine if needed
CSF200 µLLow-bind tubesFreeze immediately at −80 °CDry iceLimited volume protocols available
Tissue (animal/plant)30–100 mgCryovialsSnap-freeze; ship on dry iceDry iceRecord wet weight; avoid thawing
Cell pellets≥1×10^6 cellsLow-bind tubesWash cold PBS, snap-freezeDry iceProvide cell count and viability
Culture media1–2 mLPolypropyleneClarify, aliquot, freezeDry iceNote serum supplementation
Leaf/disc (plant)50–100 mgFoil-wrapped cryovialsDark, snap-freezeDry iceProtect from light and heat

If preservatives are restricted, we will propose a redox-safe alternative during intake.

FAQs for Oxylipins Analysis Service

What is the difference between oxylipins and eicosanoids?

Eicosanoids are a subset of oxylipins derived from arachidonic acid; oxylipins also include products from linoleic, α-linolenic, EPA, and DHA, plus plant jasmonates.

Can you separate isomeric species reliably?

Yes. We use optimized C18 gradients and multiple MRM transitions or PRM ion-ratio criteria to resolve key regio- and stereoisomers.

How do you prevent artificial oxidation during prep?

We add antioxidants and chelators, work on ice, minimize light exposure, and shorten processing time.

Do you measure conjugated forms?

We can include selected glucuronides or sulfates if standards and method conditions are defined for your matrix.

Is cross-species analysis supported?

Yes. Methods are available for human, animal, and plant matrices with matrix-matched calibration or surrogates.

How are data normalized for comparison?

We apply isotope dilution, QC-based drift correction, and optional normalization to protein, tissue weight, creatinine, or cell number.

Which analytical platform is most reliable for oxylipins—ELISA, GC-MS, or LC-MS/MS?

MS-based methods (LC-MS/MS or GC-MS) provide higher specificity and accuracy than immunoassays, which can suffer from cross-reactivity among isomers; LC-MS/MS is widely used for targeted mediator panels, while GC-MS remains a robust option for select classes such as F2-isoprostanes.

Do I need high-resolution MS for identity confirmation?

Targeted LC-MS/MS with multiple transitions can quantify most mediators; for low-abundance species (e.g., SPMs) or where isomeric interferences are possible, high-resolution PRM/HRAM overlays and ion-ratio criteria strengthen identification confidence.

Do I need to measure free and esterified oxylipins?

Free oxylipins reflect immediate signaling, whereas the esterified pool is abundant and can serve as a reservoir; many studies underreport the esterified fraction, so your choice should follow the biological question and matrix feasibility.

What does the EET/DHET ratio tell me?

The EET:DHET ratio (or specific 14,15-EET:14,15-DHET) is commonly used as a functional readout of soluble epoxide hydrolase activity and epoxy–diol turnover in vivo.

Publications

* Our services can only be used for research purposes and Not for clinical use.

Services:


Online Inquiry

CONTACT US

Services

Resource

  • USA
  • Tel:
  • Address:
  • Email:
  • Fax:
  • Germany
  • Address:
Copyright © 2025 Creative Proteomics. All rights reserved.