Omega-3 Fatty Acids Analysis Service

What is Omega-3 Fatty Acids?

Omega-3 fatty acids are a class of essential polyunsaturated fatty acids (PUFAs) characterized by the presence of a double bond at the third carbon atom from the methyl end of the fatty acid chain. These molecules are widely recognized for their structural and functional roles in biological membranes, as well as their significance in nutritional formulations and industrial lipid applications. The most commonly studied omega-3 fatty acids include:

• Alpha-linolenic acid (ALA; C18:3n-3) – a plant-derived essential fatty acid
• Eicosapentaenoic acid (EPA; C20:5n-3) – prevalent in marine oils
• Docosahexaenoic acid (DHA; C22:6n-3) – important in cellular lipid composition
• Docosapentaenoic acid (DPA; C22:5n-3) – an intermediate in metabolic conversion

These compounds are not synthesized efficiently by humans or animals and must be obtained through dietary sources such as fish oil, algal oil, flaxseed, and fortified foods. They are widely monitored and optimized in product formulations across the functional food, nutraceutical, agriculture, and cosmetic sectors.

Why Analyze Omega-3 Fatty Acids?

Analyzing omega-3 fatty acids is crucial for product development, quality assurance, and research purposes. Accurate analysis supports various applications:

• Formulation optimization: Ensures balanced omega-3 content in food, nutraceutical, and cosmetic products.
• Quality control: Confirms the consistency and purity of omega-3 in supplements and other products.
• Research and development: Facilitates the study of omega-3's role in metabolic pathways and its impact on health.
• Process optimization: Enhances extraction methods and refines production processes for raw materials.

Omega-3 analysis ensures product integrity, supports product development, and offers valuable insights for further research.

Omega-3 Fatty Acids Analysis Service Offered by Creative Proteomics

Quantification of Omega-3 Fatty Acids

• EPA, DHA, ALA, DPA, and other omega-3s are accurately quantified using LC-MS/MS and GC-MS techniques.
• Absolute quantification with internal standards ensures precise and reproducible results across multiple sample types.

Comprehensive Lipid Profiling

• Full lipidomic analysis including omega-3 fatty acids and their derivatives.
• Detection of key omega-6/omega-3 ratios to assess nutritional balance.
• Metabolites like resolvins and protectins are also identified to explore functional effects.

Isomer Profiling and Differentiation

• Analysis of DHA/EPA isomers and differentiation of omega-3 variants.
• Detection of Saturated and unsaturated isomers for in-depth product characterization.

Metabolomic Pathway Analysis

• Pathway mapping of omega-3 fatty acids and their metabolites through integrated bioinformatics tools.
• Identification of key pathways involved in lipid metabolism and inflammation resolution.

Matrix-Specific Extraction and Analysis

Customized sample extraction methods for a wide variety of matrices, including:

• Fish oils, algae oils, and plant-based sources
• Plasma, serum, tissues, and other biological samples
• Nutritional products and fortified foods
• Fermentation and production-derived samples

Stability and Shelf-Life Testing

Oxidative stability testing of omega-3-rich products. Long-term storage and shelf-life studies to monitor degradation and quality control.

Custom Analytical Services

Development of custom lipidomic panels tailored to specific research or product needs. Targeted analysis of additional fatty acids and related metabolites upon request.

High-Throughput Screening

Support for high-throughput analysis, enabling rapid screening of large sample sets. Automated sample preparation and LC-MS/MS workflows provide quick and reliable results.

List of Detected Omega-3 Fatty Acids and Related Metabolites

Advantages of Omega-3 Fatty Acids Assay

• Ultra-high sensitivity, with detection limits as low as 0.5 ng/mL, enables trace-level quantification of EPA, DHA, and related metabolites in complex matrices.
• Extensive compound coverage, detecting over 50 omega-3-related fatty acids and metabolic intermediates across multiple biosynthetic pathways.
• Exceptional reproducibility, with intra- and inter-batch coefficient of variation (CV) consistently maintained below 10% using isotope-labeled internal standards.
• Accelerated turnaround time, delivering results within 14–42 business days while supporting high-throughput capacity of up to 200 samples per day.
• Scalable analytical platform, suitable for both small-batch product evaluation and large-cohort research studies, supported by automated sample preparation and LC-MS workflows.
• Matrix versatility, accommodating a wide range of sample types including fish oils, microalgae, fortified foods, plasma, serum, tissues, and fermentation broths.
• Robust quantification range, from 0.5 ng/mL to over 100 µg/mL, meeting the needs of both trace detection and bulk compositional analysis.
• Comprehensive pathway interpretation, with detected metabolites mapped to KEGG, HMDB, and LIPID MAPS databases for full lipid pathway contextualization.
• Customized reporting formats, including concentration tables, metabolic pathway visualizations, and statistical summaries tailored to client-specific research or development objectives.

Workflow for Omega-3 Fatty Acids Analysis Service

Technology Platform for Omega-3 Fatty Acids Analysis Service

LC-MS/MS (Thermo Scientific Q Exactive™ Orbitrap) – For high-resolution quantification.

Agilent 6545XT AdvanceBio Q-TOF – For targeted lipidomics and fragmentation profiling.

GC-MS (Agilent 7890B-5977B) – For volatile and derivatized fatty acids.

NMR Spectroscopy (Bruker Avance III 600 MHz) – For absolute structural confirmation.

Thermo Fisher Q Exactive (Figure from Thermo Fisher)

Agilent 7890B-5977B (Figure from Agilent)

Bruker Avance III 600 MHz (Fig from Bruker)

Sample Requirements for Omega-3 Fatty Acids Analysis Service

Omega-3 Fatty Acids Analysis Service Case Study

Title: Omega-3 Polyunsaturated Fatty Acids Decrease Aortic Valve Disease Through the Resolvin E1 and ChemR23 Axis

Journal: Circulation

Published: 2020

• Background
• Materials & Methods
• Results
• Reference

Aortic valve stenosis (AVS) is the most common valvular heart disease, characterized by progressive narrowing of the aortic valve due to leaflet thickening and calcification. It significantly increases the risk of heart failure and cardiovascular mortality. Traditional treatments fail to halt disease progression, leaving valve replacement as the only option. Omega-3 polyunsaturated fatty acids (n-3 PUFAs), particularly EPA and DHA, have demonstrated cardiovascular benefits due to their anti-inflammatory and proresolving properties, which are mediated through specialized proresolving mediators (SPMs) like resolvin E1 (RvE1). This study investigates the role of RvE1 and its receptor ChemR23 in reducing AVS progression.

The study analyzed tricuspid aortic valves from 96 patients undergoing valve replacement surgery, with full ethical approval and informed consent. Lipidomic profiling was performed using liquid chromatography–tandem mass spectrometry to quantify specialized proresolving mediators. For mechanistic studies, genetically modified mice—including Fat-1 transgenic, Apoe−/−, ChemR23−/−, and their combinations—were used. Statistical analyses involved t-tests, ANOVAs, and mixed-effects models, with significance set at P<0.05. All procedures followed ethical standards for human and animal research.

n-3 PUFA Content in Human Aortic Valves

Noncalcified aortic valve tissue had significantly higher HS-omega-3 index compared to calcified regions. Slower AVS progression was associated with a higher HS-omega-3 index, though not statistically significant.

Lipid Mediators in Aortic Valves

n-3 PUFA-derived SPMs (RvE1 and RvD3) were lower in calcified valves. In contrast, the pro-inflammatory leukotriene B4 from n-6 PUFA was higher in calcified tissue.

RvE1 Receptor (ChemR23) in Aortic Valves

ChemR23, the RvE1 receptor, was predominantly expressed in human aortic valves and was associated with reduced calcification in vitro.

Fat-1tg×Apoe−/− Mice

These mice, with endogenous n-3 PUFA production, showed lower transaortic velocity, increased cusp separation, and reduced valve calcification compared to controls.

ChemR23 as a Key Mediator

ChemR23 deletion in Apoe−/− mice reversed the benefits of n-3 PUFAs, leading to increased Vmax, reduced cusp separation, and increased leaflet calcification.

Inflammation and Macrophage Polarization

Fat-1tg promoted M2 macrophage polarization (CD206, arginase-1) in a ChemR23-dependent manner, with no significant change in M1 markers.

Human Data Correlation

ChemR23 expression in human valves correlated with M2 macrophage markers, particularly in both calcified and noncalcified regions.

ChemR23 Deletion in Valve Injury Model

ChemR23−/− mice exhibited worse outcomes after aortic valve injury, including increased leaflet thickening and higher Vmax.

HS-omega-3 index is decreased in calcified aortic valve tissue and is lower in noncalcified valve tissue of aortic valve stenosis (AVS) fast progressors.

Omega-3 polyunsaturated fatty acids (PUFAs) are higher and n-6 PUFAs are lower in valve leaflets of Fat-1tg×Apoe−/− compared with Apoe−/− mice.

Reference

1. Artiach, Gonzalo, et al. "Omega-3 polyunsaturated fatty acids decrease aortic valve disease through the resolvin E1 and ChemR23 axis." Circulation 142.8 (2020): 776-789. https://doi.org/10.1161/CIRCULATIONAHA.119.041868.

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