Lipoxygenase Metabolite Profiling Service

Tailored Lipoxygenase Metabolite Profiling to Meet Complex Research Demands

Lipoxygenase (LOX) enzymes are central to the oxidative metabolism of polyunsaturated fatty acids, giving rise to a diverse range of bioactive lipid mediators such as hydroxy fatty acids, leukotrienes, lipoxins, and specialized pro-resolving mediators (SPMs). These metabolites play pivotal roles in cellular signaling, immune response modulation, inflammation resolution, and oxidative stress. Accurate profiling of LOX-derived metabolites is therefore essential for researchers investigating lipid signaling pathways, inflammation mechanisms, and redox biology.

At Creative Proteomics, our lipoxygenase metabolite profiling service is engineered to support lipidomics studies across a wide spectrum of biological samples and experimental models. Using high-sensitivity LC-MS/MS platforms and proprietary sample preparation protocols, we deliver reliable identification and absolute or relative quantification of a broad panel of LOX-derived metabolites with exceptional analytical specificity.

What Clients Typically Need?

Mechanistic Dissection of LOX Pathways

Many clients aim to map the specific activity of LOX isoforms (e.g., 5-LOX, 12-LOX, 15-LOX) under physiological or experimental conditions. We support this by delivering isomer-specific quantification of key metabolites such as:

• 5-HETE / 5-oxo-ETE (5-LOX pathway)
• 12-HETE (platelet-type 12-LOX)
• 15(S)-HETE / 15(R)-HETE (15-LOX-1 vs 15-LOX-2)

This allows clients to trace isoform-selective activity and investigate enzyme regulation.

Comprehensive Lipid Mediator Panel for Inflammation Research

Researchers studying inflammation, immune signaling, or resolution processes often require in-depth profiling of both pro-inflammatory and pro-resolving mediators derived from arachidonic acid (AA), docosahexaenoic acid (DHA), or eicosapentaenoic acid (EPA). Our expanded LOX panel supports:

• Detection of leukotrienes and lipoxins
• Analysis of SPMs such as resolvins and protectins
• Cross-comparison of COX-, LOX-, and CYP-derived mediators (optional add-on)

Oxidative Stress and Redox Biology Investigations

LOX-derived lipid peroxides and hydroxy fatty acids serve as sensitive markers of non-enzymatic lipid oxidation and oxidative cell injury. We provide quantitative data on oxidized linoleic acid metabolites (e.g., 9-HODE, 13-HODE) to help assess:

• Redox balance in disease models
• Impact of antioxidant treatments
• Lipid peroxidation status in response to stimuli

Nutritional, Metabolic, and Diet Intervention Studies

Clients investigating the effects of dietary polyunsaturated fatty acids (PUFAs) or supplements often need to monitor changes in lipid mediator profiles. We enable:

• Longitudinal tracking of LOX metabolites in blood or tissues
• Profiling of EPA/DHA-derived hydroxy metabolites (e.g., 17-HDHA, 14-HDHA)
• Integration with metabolic health markers for pathway correlation

Drug Discovery and Functional Screening

For pharmaceutical clients and screening labs, our service supports:

• Profiling of lipid mediators in response to LOX inhibitors or enzyme modulators
• Off-target analysis of compound effects on AA/DHA oxidation
• Dose-response characterization of small molecule activity on lipid mediator production

Lipoxygenase-Derived Metabolite Panel

Key Features of Our Lipoxygenase Metabolite Profiling Service

• High Sensitivity: Detection limits down to ≤10 pg/mL for key LOX metabolites.
• Broad Coverage: Simultaneous quantification of 40+ lipid mediators, including HETEs, leukotrienes, lipoxins, resolvins.
• Stereoisomer Resolution: Baseline separation of S-/R-HETEs with<0.05 min retention time difference.
• Quantitative Precision: Intra- and inter-run CVs<12% across replicates.
• Dynamic Range: Linear quantification over 5–6 orders of magnitude (0.01–1000 ng/mL).
• Small Sample Volume: Requires only ≥50 µL plasma or ≥10 mg tissue per replicate.
• Internal Standards: Use of 10–15 isotope-labeled compounds for accurate absolute quantification.
• Optional Customization: Add up to 20 additional analytes with minimal method adjustment.

Methods and Instrumentation for Lipoxygenase Metabolites Analysis

System Component	Model	Key Parameters
UPLC System	Waters ACQUITY UPLC	C18 reversed-phase column (2.1 × 100 mm, 1.7 μm); Column temp: 40°C; Flow: 0.3 mL/min
Mass Spectrometer	SCIEX QTRAP® 6500+ Triple Quadrupole	Electrospray Ionization (ESI), Negative Mode; Scheduled MRM acquisition
LC Gradient	Water (0.1% formic acid) / Acetonitrile (0.1% formic acid)	Gradient elution optimized for LOX-derived eicosanoids and docosanoids
Injection Volume	–	Typically 5–10 μL depending on sample matrix
MS Parameters	–	Optimized declustering potential, collision energy, and entrance potential per analyte

SCIEX Triple Quad™ 6500+ (Figure from Sciex)

Waters ACQUITY UPLC System (Figure from Waters)

Our Lipoxygenase Metabolite Assay Workflow

Sample Requirements for Lipoxygenase Metabolites Analysis Service

Demo Results

Representative LC-MS/MS chromatograms of LOX metabolites.
(A) Baseline separation of 15(S)-HETE and 15(R)-HETE in plasma. (B) Overlaid chromatograms of 5-HETE, 12-HETE, and 15-HETE demonstrating retention time differences.

Multi-omics statistical analysis of LOX metabolites. (A) Heatmap of LOX metabolite levels across sample groups. (B) PCA score plot showing group separation based on LOX profiles.

FAQ of Lipoxygenase Metabolite Profiling Service

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Publications

Here are some of the metabolomics-related papers published by our clients: