FAHFAs Analysis Service

What Are FAHFAs?

Fatty acid esters of hydroxy fatty acids (FAHFAs) are a unique class of endogenous lipids formed through the esterification of a hydroxy fatty acid and a long-chain fatty acid. Unlike conventional lipids, FAHFAs exhibit regioisomeric diversity, meaning their biological function varies depending on the position of the ester linkage—such as 9-PAHSA or 5-OAHSA.

These compounds were first recognized for their ability to modulate glucose metabolism, lipid homeostasis, and immune signaling, making them an important focus in modern lipidomics research. Structurally, FAHFAs combine hydrophobic and hydroxyl-functional domains, contributing to their stability within biological membranes and their interaction with lipid-metabolizing enzymes.

In biochemical studies, FAHFAs are now viewed as regulatory lipids that bridge metabolic and signaling pathways. Their abundance and isomeric patterns can indicate alterations in cellular lipid remodeling, oxidative stress response, or energy regulation—offering valuable insight into lipid metabolism under physiological or environmental change.

FAHFAs Analysis Solutions at Creative Proteomics

Targeted FAHFA Profiling

Quantify prioritized families and regioisomers (e.g., PAHSAs, OAHFAs, SAHFAs) with internal-standard calibration.

Isomer-Resolved Identification

Separate and confirm positional isomers using optimized reverse-phase LC and diagnostic MS/MS fragments.

Untargeted Discovery Screening

Flag unexpected FAHFA species and related hydroxy-lipid esters for follow-up targeted verification.

Absolute Quantification

Apply isotope-labeled analogs for concentration reporting in serum, plasma, tissues, cell pellets, and formulated materials.

Structural Confirmation

Validate ester linkage and hydroxy position via fragmentation mapping and retention-time orthogonality.

Comparative Lipidomics

Assess group differences with normalized peak areas, effect-direction summaries, and optional pathway context.

Full Analyte List — FAHFAs We Detect

The list below shows representative families and regioisomers commonly included in our panels. Final targets can be expanded or customized to your matrix and study goals.

How Creative Proteomics Provides FAHFAs Assay?

What Methods are Used for Our FAHFAs Analysis?

Our FAHFA assays combine high-resolution discovery and targeted quantification platforms to ensure structural confidence and absolute accuracy.

Instrumentation

• Thermo Scientific Q Exactive HF-X Orbitrap — Provides full-scan coverage at 120,000 resolution with mass accuracy within ±2 ppm, ideal for detecting isomeric and low-abundance FAHFAs.
• AB Sciex 6500+ Triple Quadrupole — Operates in scheduled MRM mode, ensuring high selectivity and linear quantification across a dynamic range of five orders of magnitude.
• Agilent 1290 Infinity II UHPLC System — Delivers stable, reproducible LC separation with retention time precision <0.1% RSD.

Chromatographic Conditions

• Column: C30 reverse-phase, 2.1 × 100 mm, 3 µm for enhanced shape selectivity.
• Mobile Phase: Gradient of acetonitrile/isopropanol/water with 10 mM ammonium formate and 0.1% formic acid.
• Flow Rate: 0.3 mL/min; Column Temp: 40 °C.

Detection Mode & Parameters

• ESI source in positive/negative polarity switching for comprehensive ion coverage.
• Collision energies optimized (20–35 eV) for diagnostic MS/MS fragment ions confirming hydroxy and ester positions.
• Internal standardization with deuterated PAHSAs and OAHFAs supports calibration curve linearity (R² ≥ 0.995).

This dual-platform strategy enables both comprehensive FAHFA discovery and precise quantitative validation, ensuring publication-grade data for research across metabolism, nutrition, and lipid biochemistry.

SCIEX Triple Quad™ 6500+ (Figure from Sciex)

Q Exactive HF-X MS

Agilent 1260 Infinity II HPLC (Figure from Agilent)

FAHFAs Analysis Service: Results and Data Analysis

Standard Deliverables

• Quantitative Result Tables — Absolute or relative concentrations (e.g., ng/mL, pmol/mg) of all detected FAHFA species, with isomer-specific values (e.g., 5-, 9-, 12-PAHSA).
• FAHFA Class Summary Reports — Aggregated abundance of major FAHFA families (PAHSA, OAHFA, SAHFA, DHAHFA, EPAHFA) for comparative interpretation across matrices.
• Normalized Data Matrices — Internal-standard–corrected and sample-normalized data formatted for import into statistical tools such as MetaboAnalyst.
• Full Raw Data Files — Provided in Thermo .raw and open .mzML formats to ensure transparency and reproducibility.
• Quality Control Summary — Includes internal standard recovery, calibration linearity (R² ≥ 0.995), and RSD% for analytical repeatability.
• Annotated LC–MS/MS Chromatograms — Key chromatographic peaks and MS/MS spectra confirming FAHFA identity and retention stability.

Isomer-Resolved LC–MS/MS Chromatogram with Annotated Fragmentation Spectra

Diagnostic Fragmentation Map for FAHFA Structural Identification

What Our FAHFAs Analysis Used For

Metabolic Research

Map FAHFA isomer patterns to lipid and energy regulation.

Nutritional Science

Track diet-induced shifts in ω-3/ω-6 FAHFA families.

Redox Biology

Assess oxidation-sensitive FAHFAs under stress conditions.

Pharmacology & Screening

Evaluate compound effects on FAHFA biosynthesis and turnover.

Cellular Differentiation Studies

Profile FAHFAs during lineage commitment and remodeling

Microbiome–Lipid Interaction Studies

Explore how gut or skin microbiota influence FAHFA

Sample Requirements for FAHFAs Analysis Solutions

FAQs for FAHFAs Analysis Service