Precision Analysis for Unsaturated Fatty Acids

What Are Unsaturated Fatty Acids?

Unsaturated fatty acids—commonly referred to as unsaturated fats—are a type of fat that remains liquid at room temperature. Unlike saturated fats, these molecules contain at least one double bond in their carbon chain:

If there's one double bond, it's a monounsaturated fat (e.g., oleic acid).

If there are multiple double bonds, it's a polyunsaturated fat (e.g., omega-3 and omega-6 fatty acids).

These structural differences affect both their biological behaviour and health impact. For instance, unsaturated fats yield less energy during cellular metabolism compared to saturated fats. However, they are also more susceptible to oxidation, especially those with multiple double bonds—a process that can be slowed by antioxidants like vitamin E.

You'll find natural sources of unsaturated fats in:

Avocados and nuts

Plant-based oils such as olive, canola, and soybean oil

Animal products, which often contain both saturated and unsaturated fats

Unsaturated vs. Saturated Fats: What the Science Says

Saturated fats, which have no double bonds, tend to be solid at room temperature and are found in animal fats, butter, and some plant oils. They are known to raise LDL cholesterol levels, increasing the risk of heart disease. In contrast, unsaturated fats—especially PUFAs and MUFAs—can improve lipid profiles, reduce inflammation, and support healthy blood pressure.

Dietary studies, including those examining the Mediterranean diet, strongly support the health benefits of replacing saturated fats with unsaturated fats. As such, many regulatory bodies recommend increasing the proportion of UFAs in daily intake.

Why Unsaturated Fatty Acid Analysis Matters

Understanding the presence, concentration, and behavior of unsaturated fatty acids is essential in various industries. In the food industry, accurate analysis ensures compliance with nutritional labeling standards and helps identify adulteration in products like olive oil or margarine. In the pharmaceutical sector, UFAs are studied for their role in drug absorption and delivery, especially in lipid-based formulations.

Moreover, researchers in academic settings use unsaturated fatty acid analysis to explore their involvement in metabolic pathways and diseases. The increasing consumer awareness regarding the benefits of healthy fats is also driving demand in nutraceutical and functional food sectors, where rigorous testing is required to verify label claims and efficacy.

The Role of UFA Metabolism in Human Health

Unsaturated fatty acids contribute to multiple metabolic functions. Omega-3 fatty acids like EPA and DHA are known to reduce triglyceride levels and suppress inflammation, while omega-6 fatty acids like linoleic acid help with skin health and brain function. These fatty acids are metabolized into signaling molecules such as prostaglandins and leukotrienes, which play roles in immune regulation.

An imbalance in the intake of omega-6 and omega-3 fatty acids can lead to chronic inflammation. Therefore, it's vital to monitor both dietary intake and metabolic utilization, especially for patients with metabolic disorders such as obesity, diabetes, or cardiovascular disease.

How Are Unsaturated Fatty Acids Analyzed?

Several sophisticated analytical techniques are employed to analyze unsaturated fatty acids. The most commonly used methods include:

• High-Performance Liquid Chromatography (HPLC): A preferred method due to its precision and ability to differentiate between isomers.
• Gas Chromatography-Mass Spectrometry (GC-MS): Often used for the analysis of fatty acid methyl esters (FAMEs), offering high sensitivity and specificity.
• Liquid Chromatography-Mass Spectrometry (LC-MS): Ideal for in-depth metabolic studies and flux analysis of fatty acids in complex biological samples.

Creative Proteomics provides a comprehensive suite of services that utilize these technologies for high-quality results.

Creative Proteomics' UFA Analysis Service

At the heart of our Unsaturated Fatty Acids Analysis Service is a robust LC-MS/MS platform, tailored to meet the precision demands of researchers in drug development, nutrition science, and academic labs.

We use isotope-labelled internal standards alongside external calibration curves to ensure every data point is both accurate and reproducible. This dual-approach quantification offers:

• High sensitivity, ideal for low-abundance analytes
• Low detection limits to capture subtle biological shifts
• A broad dynamic range for profiling diverse sample types

We can simultaneously quantify 11 different unsaturated fatty acids in a single run—maximising efficiency without compromising data quality.

Whether you're studying lipid metabolism or screening biomarkers for disease, our platform delivers the reliability and throughput you need.

Related services include:

Straight Chain Fatty Acids Analysis Service

Saturated Fatty Acids Analysis Service

Total Fatty Acids Analysis Service

Key Features of Our Unsaturated Fatty Acids Analysis Service

Experienced Analytical Team: Our scientists bring years of hands-on expertise in lipid profiling, ensuring accurate interpretation and high-quality data delivery.

• High-Throughput Performance: Designed for scalability, our platform handles large sample batches efficiently—ideal for time-sensitive or high-volume projects.
• Flexible and Fast: Choose from customizable service packages with rapid turnaround times to meet your unique research timelines.
• Regulatory & Publication Support: From FDA-compliant documentation to figures for peer-reviewed journals, we've got your reporting needs covered.
• Exceptional Separation Efficiency: Leveraging subtle differences in partition coefficients, we achieve precise separation—even for structurally similar isomers.
• Ultra-Sensitive Detection: Using a flame ionization detector (FID), we reach detection limits in the microgram range—perfect for trace-level fatty acid quantification.
• Rapid Chromatographic Analysis: Optimized columns and run conditions enable fast analysis times—from just a few minutes to under an hour—supporting high-throughput demands.
• Reliable Quantitative Accuracy: By combining external calibration with isotope-labelled internal standards, we deliver consistent, reproducible data with high repeatability.
• Broad Application Scope: Whether you're studying common fatty acids or niche lipid species, our method adapts seamlessly to diverse sample types across multiple industries.

**Workflow:**

1. **Sample Reception & Validation** – Each sample undergoes quality checks.

2. **Lipid Extraction** – Using optimized protocols to ensure yield and purity.

3. **Chromatographic Analysis** – Employing HPLC or GC-MS as needed.

4. **Data Interpretation** – Detailed analysis with visual reports.

5. **Consultation & Delivery** – Optional expert consultation to guide your next steps.

Platform

Detection Method: Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS)

Instrumentation: Ultra-High Performance Liquid Chromatograph (UHPLC) and tandem mass spectrometer

Ion Source & Scan Mode: Electrospray Ionization (ESI) with Multiple Reaction Monitoring (MRM)

To enhance sensitivity, we apply time-segmented monitoring based on each compound's retention time—enabling more accurate detection of low-abundance fatty acids.

Core Technologies and Instruments:

Agilent 1290-6470 (image source: agilent.com)

Sciex QTRAP 6500+ (image source: Sciex QTRAP 6500+)

Applications Across Key Industries

Our unsaturated fatty acid profiling service is applied across multiple industries:

- **Food & Beverage**: Ensures accurate nutritional content, detects adulteration, validates omega-3 and omega-6 claims.

- **Pharmaceuticals**: Analyzes lipid-based drug carriers and tracks UFA metabolism during preclinical trials.

- **Nutraceuticals**: Supports product development of supplements containing fish oil, flaxseed oil, or other UFA-rich ingredients.

- **Cosmetics**: Evaluates lipid ingredients in creams and lotions for moisturizing efficacy and safety.

Metabolist Coverage

Table: Targeted Unsaturated Fatty Acids Covered in Our Analysis

Sample Requirements

Deliverables

You'll receive a comprehensive technical report that includes:

• Full experimental protocol and GC-MS parameters
• Quantified analytes in μM or μg/mg (tissue)
• Coefficients of variation (CV) typically<10%
• Complete fatty acid data including:
• Name, abbreviation, molecular weight
• Formula and CAS number

Case Study: Client Success Story

Fatty Acid Profiling of Nutrient-Enriched Eggs from Pasture-Raised Hens

• Background
• Methods
• Key Findings
• Conclusion

In a 2022 study published in Foods (DOI: 10.3390/foods11213404), researchers from Michigan State University evaluated how supplementing pasture-raised hens with grass-fed beef (GFB) suet and liver—instead of standard corn and soy feed—affects the nutrient composition of egg yolks. The study aimed to characterize differences in fatty acid profiles, antioxidants, vitamins, and minerals across eggs from three hen groups:

PBB: hens fed a corn- and soy-free diet with GFB by-products

PCS: hens fed a standard corn/soy diet

CFC: commercial cage-free eggs from retail sources

To assess the fatty acid composition of egg yolks, layer hen feeds, and beef by-products, the research team employed gas chromatography–mass spectrometry (GC-MS).

Creative Proteomics provided expert analytical support, performing targeted unsaturated fatty acid analysis. Fatty acids were extracted using a microwave-assisted method, then converted to fatty acid methyl esters (FAMEs) for GC-MS separation and identification.

The analysis focused on key lipid markers, including:

• Omega-3 fatty acids (e.g., ALA, DPA, DHA)
• Omega-6 fatty acids (e.g., linoleic acid)
• Conjugated linoleic acid (CLA)
• Odd- and branched-chain fatty acids (OCFAs, BCFAs)

• PBB eggs had significantly lower omega-6 PUFA and higher concentrations of CLA, OCFAs, and BCFAs than commercial cage-free eggs.
• Both PBB and PCS egg groups (pasture-raised) showed increased omega-3 PUFA content and a lower n-6:n-3 ratio, aligning more closely with human nutritional guidelines.
• Despite the inclusion of GFB suet (a saturated fat source), total saturated fatty acid content did not increase, suggesting a favorable lipid shift.

Creative Proteomics' unsaturated fatty acid profiling service was essential in uncovering these compositional changes—enabling the precise quantification of lipid classes associated with improved egg nutrition and sustainable feed practices.

Table. Fatty acid profile of the egg yolks (% of total fatty acids)

Fatty Acid	PBB Eggs	PCS Eggs	CFC Eggs	p-Value 2
10:0	0.02 ± 0.00	0.02 ± 0.01	0.02 ± 0.00	0.482
12:0	0.01 ± 0.00	0.01 ± 0.00	0.01 ± 0.00	0.442
14:0	0.56 ± 0.13 a	0.29 ± 0.10 b	0.27 ± 0.08 b	<0.001
14:1	0.13 ± 0.05 a	0.03 ± 0.01 b	0.02 ± 0.01 b	<0.001
16:0	26.60 ± 1.90	28.12 ± 1.80	28.53 ± 1.64	0.177
16:1 n-7	3.24 ± 0.88 a	2.54 ± 0.63 ab	1.71 ± 0.44 b	0.005
16:1 n-7 t	0.13 ± 0.04 a	0.09 ± 0.03 b	0.08 ± 0.01 b	0.004
16:1 n-9	0.70 ± 0.16 a	0.51 ± 0.09 b	0.37 ± 0.07 b	0.001
18:0	7.43 ± 0.37	7.00 ± 1.33	7.83 ± 1.45	0.479
18:1 n-7	1.97 ± 0.29 a	1.74 ± 0.21 a	1.14 ± 0.15 b	<0.001
18:1 n-9	46.62 ± 2.16 a	39.02 ± 2.74 b	35.18 ± 3.34 b	<0.001
18:1 n-9 t	0.32 ± 0.04 a	0.19 ± 0.04 b	0.17 ± 0.01 b	<0.001
18:2 n-6	7.08 ± 1.46 c	16.19 ± 3.91 b	21.76 ± 4.63 a	<0.001
18:3 n-3	0.40 ± 0.10 ab	0.62 ± 0.27 a	0.16 ± 0.04 b	0.001
18:3 n-6	0.11 ± 0.01 b	0.15 ± 0.05 ab	0.17 ± 0.01 a	0.015
20:0	0.04 ± 0.00	0.04 ± 0.02	0.04 ± 0.00	0.882
20:1 n-9	0.34 ± 0.06 a	0.19 ± 0.07 b	0.18 ± 0.02 b	<0.001
20:2 n-6	0.07 ± 0.04 b	0.19 ± 0.05 a	0.25 ± 0.08 a	<0.001
20:3 n-6	0.13 ± 0.01	0.16 ± 0.06	0.17 ± 0.01	0.171
20:4 n-6	0.61 ± 0.14	0.65 ± 0.13	0.79 ± 0.07	0.050
20:5 n-3	< LLOD	< LLOD	< LLOD	-
22:0	< LLOD	< LLOD	< LLOD	-
22:4 n-6	0.30 ± 0.10	0.33 ± 0.21	0.16 ± 0.14	0.176
22:5 n-3	0.33 ± 0.04 a	0.19 ± 0.12 b	< LLOD c	<0.001
22:5 n-6	0.26 ± 0.05 b	0.27 ± 0.08 b	0.38 ± 0.03 a	0.006
22:6 n-3	0.77 ± 0.10 a	0.93 ± 0.23 a	0.30 ± 0.05 b	<0.001
24:0	< LLOD	< LLOD	< LLOD	-
Total SFA	35.21 ± 2.10	35.74 ± 2.27	36.88 ± 2.27	0.431
Total MUFA	53.84 ± 1.66 a	44.42 ± 2.82 b	38.92 ± 3.68 c	<0.001
Total PUFA	10.36 ± 1.65 b	19.74 ± 3.72 a	24.19 ± 4.66 a	<0.001
Total n-6	8.55 ± 1.62 c	17.94 ± 3.68 b	23.67 ± 4.59 a	<0.001
Total n-3	1.50 ± 0.12 a	1.74 ± 0.39 a	0.47 ± 0.08 b	<0.001
n-6:n-3 ratio	5.72 ± 1.12 c	10.79 ± 3.30 b	50.63 ± 4.21 a	<0.001

This case highlights how high-quality unsaturated fatty acid analysis supports evidence-based improvements in functional foods. With Creative Proteomics' GC-MS expertise, the study demonstrated how dietary intervention in hens can enhance egg yolk nutritional profiles, reduce agricultural waste, and align with regenerative farming goals.

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