Straight Chain Fatty Acids Analysis Service

What are Straight Chain Fatty Acids?

Straight chain fatty acids are the basic building block in the universe of lipids. They are composed of a carboxylic acid with a long and unbranched chain of carbon atoms. The number of carbons in the fatty acid varies among species. In higher organisms like mammals, fatty acids always consist of an even number of carbons, however in lower organisms like bacteria, odd number of carbon also exists. Most naturally occurring fatty acids have 4 to 26 carbons, but longer fatty acids of up to 35 carbons are also described in literature.

According to the number of carbons, straight chain fatty acids can be classified into short chain fatty acids (straight chain fatty acid, C2–C7), medium chain fatty acids (MCFA, C8–C13), long chain fatty acids (LCFA, C14–C19), and very long chain fatty acids (VLCFA, C20+). Straight chain fatty acids are either saturated or unsaturated with one or more double bonds.

Figure 1. Straight chain fatty acids: hexadecanoic acid

In cells, straight chain fatty acids play important biological roles. They have a variety functions, including being incorporated into membrane for membrane proliferation, energy storage, generation of signaling molecules, or oxidized to carbon dioxide as energy source. The levels of straight chain fatty acids are sensitive to metabolic perturbations, therefore, they can be served as good markers for many health conditions and diseases. So far, the profiling of straight chain fatty acid has lead to the discovery of many disease markers, including insulin resistance, type 2 diabetes, obesity, cardiovascular diseases, cancers, and many other diseases.

Creative Proteomics offers a comprehensive profiling of straight chain fatty acids that satisfy a variety of purposes. GC/MS platform enables more sensitive and selective assay of fatty acid measurement. GC/MS techniques serves as a powerful analytical tool for identification and quantification of fatty acids.

Straight Chain Fatty Acids Analysis in Creative Proteomics

Targeted Metabolomics Analysis of Straight Chain Fatty Acids

Measure the concentration of individual straight chain fatty acids in samples. Applications include nutritional studies, clinical diagnostics, and metabolic research. The output is an accurate quantification of straight chain fatty acids such as acetic acid, propionic acid, and butyric acid.

Straight Chain Fatty Acids Profiling

Comprehensive profiling of straight chain fatty acids in complex matrices. Applications include microbiome studies, food and beverage quality control, and environmental monitoring. The output is a detailed profile of straight chain fatty acids present in the sample, including relative abundances.

Metabolic Pathway Analysis

Investigate the metabolic pathways involving straight chain fatty acids. Applications include biochemical pathway elucidation, metabolic engineering, and disease mechanism studies. The output provides insights into the metabolic pathways and interactions involving straight chain fatty acids.

Comparative Analysis

Compare straight chain fatty acid levels across different conditions or sample types. Applications include biomarker discovery, treatment efficacy studies, and comparative microbiome research. The output is comparative data showing variations in straight chain fatty acid levels between different sample sets.

Time-Course Analysis

Monitor changes in straight chain fatty acid concentrations over time. Applications include kinetic studies, fermentation process optimization, and dynamic metabolic studies. The output is temporal data illustrating straight chain fatty acid concentration changes at various time points.

Source Identification

Identify the origin of straight chain fatty acids in samples. Applications include traceability studies, source attribution in environmental samples, and food authenticity testing. The output is the identification of straight chain fatty acid sources based on unique profiles and isotopic signatures.

Functional Role Analysis

Explore the functional roles of straight chain fatty acids in biological systems. Applications include health and disease research, cellular signaling studies, and therapeutic target identification. The output provides functional insights into the roles of straight chain fatty acids in biological processes.

Analytical Techniques for Straight Chain Fatty Acids Analysis

Gas Chromatography-Mass Spectrometry (GC-MS)

GC-MS is a highly sensitive and specific technique for the separation, identification, and quantification of SCFAs. The Agilent 7890B GC System, paired with the Agilent 5977B Mass Selective Detector (MSD), offers unparalleled performance in SCFA analysis.

Features:

• High separation efficiency
• Accurate mass detection
• Capability to handle complex mixtures

Liquid Chromatography-Mass Spectrometry (LC-MS)

LC-MS combines the physical separation capabilities of liquid chromatography with the mass analysis capabilities of mass spectrometry. The Thermo Scientific Q Exactive Orbitrap LC-MS/MS is ideal for comprehensive profiling of SCFAs, providing high resolution and precision.

Features:

• High-resolution mass spectrometry
• Versatile sample compatibility
• Detailed molecular characterization

Nuclear Magnetic Resonance (NMR) Spectroscopy

NMR spectroscopy is a non-destructive technique that provides detailed structural information about SCFAs. The Bruker Avance III 600 MHz NMR Spectrometer is used for high-resolution NMR analysis, enabling precise identification and quantification of SCFAs.

Features:

• High sensitivity and resolution
• Detailed structural elucidation
• Reproducibility

High-Performance Liquid Chromatography (HPLC)

HPLC is a powerful technique for the separation and quantification of SCFAs. The Waters ACQUITY UPLC H-Class System provides high resolution and reproducibility, making it ideal for a wide range of sample types.

Features:

• High separation efficiency
• Robust and reliable performance
• Versatility in sample analysis

List of Straight Chain Fatty Acids Analysis

Sample Requirements for Straight Chain Fatty Acids Analysis

Important Considerations:

• Storage: Samples should be stored at appropriate temperatures to maintain lipid integrity.
• Preparation: Proper extraction methods must be used to avoid contamination or loss of fatty acids.
• Documentation: Detailed sample information (e.g., source, processing, additives) is crucial for accurate analysis.
• Handling: Minimize exposure to light, heat, and oxygen during transportation and storage.

Efficacy of dietary odd-chain saturated fatty acid pentadecanoic acid parallels broad associated health benefits in humans: could it be essential

Journal: Scientific reports

Published: 2020

Background

The study focuses on pentadecanoic acid (C15:0), a saturated fatty acid, hypothesized to act as a ligand for peroxisome proliferator-activated receptors (PPARs) and potentially possess anti-inflammatory and antifibrotic properties. This hypothesis was based on its structural similarity to other known PPAR ligands and its potential benefits in cellular systems related to inflammation and fibrosis.

Materials & Methods

Free Fatty Acids

Synthetic saturated fatty acids, including pentadecanoic acid (C15:0) and heptadecanoic acid (C17:0), were sourced from Millipore Sigma and used in the studies.

Cell-based PPAR Agonist Assays

Cell-based assays using CHO cell lines were employed to assess the agonist activities of C15:0 on PPARα, PPARδ, and PPARγ receptors. Assays were conducted using the Eurofins/DiscoverX PathHunter NHRscan panel, comparing C15:0 against known PPAR agonists.

Off-target Pharmacology

The Eurofins/DiscoverX SafetyScan47 panel was utilized to evaluate potential off-target effects of C15:0 across various assays targeting 47 genes and pathways related to compound safety and mechanisms of action.

Mitochondrial Reactive Oxygen Species (ROS) Assays

An in-house assay using HepG2 cells measured mitochondrial ROS production in response to varying concentrations of C15:0, utilizing MitoSOX Red as a probe and analyzed by flow cytometry.

Human Cell Phenotypic Profiling

Primary human cell systems from the BioMAP Diversity PLUS panel were used to assess anti-inflammatory and antifibrotic activities of C15:0. Biomarker activities across 148 endpoints were quantitatively measured to evaluate efficacy.

Lipidomic Analysis

Lipidomic profiling was performed using liquid chromatography coupled with mass spectrometry (LC-MS) to characterize changes in lipid composition induced by C15:0 treatment. Lipid extracts from treated cells and tissues were analyzed to identify alterations in lipid species and pathways affected by C15:0.

Pharmacokinetics Study

A 24-hour pharmacokinetic study in Sprague Dawley rats administered C15:0 orally determined serum concentrations using deuterated forms of C15:0 and C17:0, analyzed by capillary gas chromatography/mass spectrometry.

14-day Toxicology Study

Sprague Dawley rats were orally dosed with C15:0 daily for 14 days to evaluate safety at varying doses. Clinical observations, body weight measurements, clinical chemistries, and organ histopathology were assessed.

Oral Supplement Studies

In vivo studies using high-fat diet-induced obesity models in mice and non-alcoholic steatohepatitis (NASH) models in rabbits investigated the effects of oral C15:0 supplementation on metabolic indices, inflammatory biomarkers, liver histology, and blood-based variables.

Results

PPAR Agonist Activity

The cell-based assays demonstrated that C15:0 significantly activated PPARα, PPARδ, and PPARγ receptors in a dose-dependent manner (Figure 1). Compared to controls and reference agonists, C15:0 exhibited robust agonist activity across all PPAR isoforms, particularly notable in PPARα and PPARγ activation assays. These findings suggest potent regulatory effects of C15:0 on lipid metabolism and inflammatory pathways mediated by PPAR signaling.

Mitochondrial ROS Production

Treatment with C15:0 resulted in a dose-dependent reduction in mitochondrial reactive oxygen species (ROS) production in HepG2 cells (Figure 2). The MitoSOX Red assay showed significant decreases in ROS levels compared to untreated controls, indicating potential antioxidant properties of C15:0, crucial for mitigating oxidative stress in metabolic disorders.

Lipidomic Profiling

Lipidomic analysis revealed distinct alterations in lipid composition upon C15:0 treatment. LC-MS identified significant changes in specific lipid species, including increased levels of phospholipids and decreased triglycerides in treated samples compared to controls (Figure 3). These findings suggest a regulatory role of C15:0 in lipid metabolism pathways, influencing cellular membrane dynamics and metabolic homeostasis.

Annotated dose-dependent anti-inflammatory and antifibrotic activities of saturated fatty acids (C13:0, C14:0, C15:0, and C16:0 in 20 µM) in primary human cell systems mimicking inflammation and fibrosis.

Pharmacokinetics and Safety

Pharmacokinetic studies demonstrated rapid absorption and distribution of C15:0 in rat serum following oral administration. Peak concentrations were observed within 2 hours, with a steady decline over 24 hours (Figure 4). Toxicology assessments over 14 days indicated no significant adverse effects on clinical observations, body weight, or organ histopathology, affirming the safety profile of C15:0 at therapeutic doses.

Plasma deuterated C15:0 (a), C17:0 (b), and C13:0 (c) concentrations achieved over 24 h in male Sprague Dawley rats (n = 6 total, 3 per time point between 15 min and 12 h) dosed orally once with deuterated C15:0 (35 mg/kg body weight).

alt: Plasma deuterated C15:0 (a), C17:0 (b), and C13:0 (c) concentrations over 24 hours in male Sprague Dawley rats.

Comparisons of circulating concentrations of the pro-inflammatory chemokine, monocyte chemoattractant 1 (MCP-1) (a), pro-inflammatory cytokine, interleukin 6 (IL-6) (b), glucose (c), and total cholesterol (d) between high fat diet-induced obese in vivo model supplemented with low dose daily oral C15:0 (pentadecanoic acid, 5 mg/kg) over 12 weeks and non-supplemented controls.

Reference

1. Venn-Watson, Stephanie, Richard Lumpkin, and Edward A. Dennis. "Efficacy of dietary odd-chain saturated fatty acid pentadecanoic acid parallels broad associated health benefits in humans: could it be essential?." Scientific reports 10.1 (2020): 8161.