Oxidative Stress Marker Analysis Service

What Are Oxidative Stress Markers — and Why Analyze Them?

Oxidative stress markers are measurable molecules produced when reactive oxygen/nitrogen species (ROS/RNS) interact with lipids, proteins, and nucleic acids. They capture mechanistic readouts of damage (e.g., lipid peroxidation, protein carbonylation, DNA oxidation) and defense status (e.g., GSH/GSSG ratio, enzymatic activity), enabling you to:

• Validate mechanisms in cell/tissue models and preclinical studies
• Rank compounds by pro-/anti-oxidant liability
• Optimize bioprocesses (cell culture media, feed strategies)
• Characterize materials and foods for oxidation stability
• Support safety and toxicology research with targeted, quantitative endpoints

What Problems We Solve for Our Clients

Low-abundance & unstable markers

Challenge: 8-iso-PGF2α, 8-OHdG, and 4-HNE adducts are scarce and degradation-prone.

Our approach: Isotope-dilution LC-MS/MS with scheduled MRM; immediate stabilization (NEM/BHT/DTPA); cooled, low-light handling.

Outcome: LLOQ 0.01–0.05 ng/mL, recovery 85–115%, intra-run CV ≤10%.

Isomeric interference

Challenge: F2-isoprostanes co-elute with prostaglandins; probe by-products overlap in m/z.

Our approach: UPLC (C18, 1.7 µm) for baseline separation; ion-ratio locking; HRAM confirmation when required.

Matrix effects & ion suppression

Challenge: Variable biological matrices distort response and linearity.

Our approach: SPE cleanup; matrix-matched calibration; post-column infusion mapping; internal-standard normalization.

Artifactual oxidation during handling

Challenge: Ex vivo oxidation inflates MDA/4-HNE and oxidizes GSH.

Our approach: Preservative SOPs, process blanks, and isotope-recovery monitors across the workflow.

Outcome: Blanks & carryover <20% of LLOQ; artifacts minimized and documented.

Limited sample volume with broad coverage needs

Challenge: Small inputs must support multi-analyte decisions.

Our approach: Multiplexed panels spanning lipid/protein/DNA/NO/thiol markers; microflow UPLC with optimized dwell times.

Outcome: ≥40 analytes quantified per run with inter-batch CV ≤12%.

Batch-to-batch comparability in large studies

Challenge: Instrument drift and extraction variability degrade longitudinal analyses.

Our approach: Pooled QCs every set, SST monitoring, and LOESS-based drift correction.

Outcome: Inter-batch CV ≤12% and QC pass rate ≥85%.

Cross-matrix comparability

Challenge: Differences between urine, plasma, tissue, and cell systems hinder comparisons.

Our approach: Creatinine/protein/wet-weight normalization with cross-matrix validation.

Outcome: Recovery 85–115% and consistent ratios (e.g., GSH/GSSG).

Migration from ELISA/colorimetry

Challenge: Need higher specificity and wider range without losing continuity.

Our approach: Structured bridging studies (ELISA ↔ LC-MS/MS), method transfer packs (MRM list, ranges, acceptance criteria).

Outcome: r ≥0.85 agreement and scalable quantification on LC-MS/MS.

What We Offer — Oxidative Stress Marker Analysis Services

• Targeted Oxidative Stress Panels (LC-MS/MS & GC-MS/MS): Quantitative panels across lipid peroxidation, protein oxidation, DNA/RNA oxidation, thiol redox, and nitrosative pathways.
• HRAM Confirmation & Structural ID: Orbitrap-based confirmation for critical calls, isomer resolution support, and adduct verification.
• Antioxidant & Enzymatic Readouts: TAC (FRAP/ABTS/ORAC/CUPRAC) and enzyme activities (SOD, CAT, GPx/GR) for complementary antioxidant capacity profiling.
• ROS/RNS Readouts (Cells & Lysates): Probe-based measurements (e.g., DHE oxidation to 2-hydroxyethidium; H₂O₂ assays) with method blanks and spike recoveries.
• Custom Panel Design & Method Transfer: Matrix-specific target selection, isotope-labeled internal standards, reporting units/thresholds aligned to your SOPs, and ELISA-to-LC-MS/MS bridging.

Oxidative Stress Markers & Detectable Analytes

Why Choose Our Oxidative Stress Marker Analysis Service: Key Advantages

• Quantitative sensitivity & range — Sub-ng/mL LLOQs across key classes (F2-isoprostanes, 8-OHdG, 4-HNE adducts) with 3–4 orders dynamic range and R² ≥0.995 calibration.
• High analytical specificity — Baseline isomer resolution for critical pairs and optional HRAM confirmation; multi-transition ion-ratio checks for confident IDs.
• Isotope-dilution coverage — Stable-isotope internal standards for >70% of targeted analytes to control recovery and matrix effects.
• Low input, high multiplexing — Quantify ≥40 analytes per run from small inputs (e.g., ≤50 μL plasma/serum or ~10 mg tissue) with maintained precision.
• Reproducibility at scale — Dense QC cadence and drift correction deliver inter-batch CV ≤12% and QC pass rate ≥85% in large studies.
• Artifact control — Preservative/stabilizer SOPs plus process blanks and carryover control keep blanks & carryover <20% of LLOQ.
• Transparent deliverables — Full method and data package (MRM lists, retention times, calibration/QC tables; raw/processed files on request) for audit-ready traceability.
• Configurable & extensible — Matrix-specific panels, add/remove targets, and ELISA-to-LC-MS/MS bridging (r ≥0.85) to preserve continuity with legacy datasets.

Instrument Platforms for Accurate Oxidative Stress Marker Quantification

Waters ACQUITY UPLC System (Figure from Waters)

SCIEX Triple Quad™ 6500+ (Figure from Sciex)

Agilent 1260 Infinity II HPLC (Figure from Agilent)

Agilent 6495C Triple quadrupole (Figure from Agilent)

How Our Oxidative Stress Marker Assay Works — Step-by-Step Process

Sample Requirements for Oxidative Stress Marker Assay

Deliverables: What You Receive from Our Oxidative Stress Marker Analysis

• Quantitative Results: Analyte concentrations (e.g., ng/mL, µM, or normalized units) with LLOQ/ULOQ, accuracy, precision, and recovery.
• QA/QC Package: Calibration curves, R², QC acceptance (±15%/±20% at LLOQ), blanks, carryover, and system suitability.
• Chromatographic & MS Evidence: Representative chromatograms, MRM transition lists, retention times, HRAM confirmations (when applicable).
• Study Report: Methods, sample prep details, internal standards used, normalization steps, and data dictionaries.
• Optional Analytics: Group statistics, volcano plots, heatmaps, and correlation with other endpoints.
• Data Files (on request): Raw vendor files (e.g., .wiff/.raw), processed tables (.csv/.xlsx), and instrument methods.

Calibration curve showing linear response of oxidative stress marker quantification. Excellent correlation (R² ≥ 0.995) across four orders of magnitude demonstrates robust sensitivity and accuracy.

Intra- and inter-batch precision results for low, mid, and high QC levels. All coefficients of variation (CV%) remain below 12%, well within acceptance limits.

Representative LC-MS/MS chromatogram of a target analyte and its internal standard. Clear co-elution and well-defined peaks illustrate assay specificity and signal quality.

Heatmap of multiple oxidative stress markers across different samples. Distinct concentration patterns highlight the breadth of analyte coverage and variability among samples.