Free Fatty Acids: Structure, Metabolism, Functions, and Measurement

In the realm of biochemistry, free fatty acids (FFAs) play a crucial role in numerous physiological processes. These versatile molecules serve as a significant source of energy and participate in signaling pathways, lipid synthesis, and various cellular functions. In this article, we will delve into the world of FFAs, exploring their structure, metabolism, presence in the bloodstream, functions, and measurement methods. As a leading contact research organization (CRO) company, Creative Proteomics is dedicated to helping researchers unravel the complexity of FFAs and their impact on human health.

Fig. 1 Free fatty acid receptors in health and disease.Fig. 1 Free fatty acid receptors in health and disease. (Kimura, Ikuo, et al, 2019)

What are Free Fatty Acids?

FFAs, also known as non-esterified fatty acids (NEFAs), are organic molecules consisting of a long hydrocarbon chain, typically composed of 12 to 24 carbon atoms, with a carboxylic acid group (-COOH) at one end. These fatty acids can be categorized into saturated and unsaturated forms, depending on the presence of double bonds in the carbon chain. Examples of common FFAs include palmitic acid, oleic acid, and linoleic acid.

Free Fatty Acid Structure

The structure of FFAs is characterized by a hydrophilic carboxyl group at one end and a hydrophobic hydrocarbon tail at the other. This amphipathic nature allows FFAs to interact with both aqueous and lipid environments. The length and degree of unsaturation in the carbon chain influence the physical properties and biological activities of FFAs.

Free Fatty Acid Metabolism

Free fatty acid metabolism refers to the processes involved in the breakdown and utilization of FFAs in the body. The metabolism of FFAs is a tightly regulated process that occurs primarily in the liver and adipose tissue. During periods of energy surplus, excess dietary fats or stored triglycerides are broken down into FFAs through lipolysis. The released FFAs are transported in the bloodstream, bound to albumin, to reach various target tissues where they can be utilized for energy production or stored as intracellular lipid droplets. Here's a concise overview of free fatty acid metabolism:

  • Mobilization: Stored fats in adipose tissue are hydrolyzed by the enzyme lipase, releasing FFAs into the bloodstream.
  • Transport: FFAs bind to albumin and are transported in the bloodstream to target tissues such as muscle and liver.
  • Uptake: FFAs are taken up by cells through fatty acid transporters located on the cell membrane.
  • Activation: Inside the cell, FFAs are activated to fatty acyl-CoA by an enzyme called fatty acyl-CoA synthetase, requiring ATP.
  • Beta-oxidation: Fatty acyl-CoA undergoes a series of enzymatic reactions in the mitochondria to generate acetyl-CoA, producing ATP and reducing equivalents (NADH and FADH2).
  • Citric Acid Cycle: Acetyl-CoA enters the citric acid cycle (also known as the Krebs cycle) to further generate ATP and reducing equivalents.
  • Electron Transport Chain: The reducing equivalents (NADH and FADH2) generated in beta-oxidation and the citric acid cycle enter the electron transport chain, leading to the production of ATP through oxidative phosphorylation.
  • Energy utilization: ATP generated during beta-oxidation, the citric acid cycle, and oxidative phosphorylation is used for cellular energy needs or stored for future use.
  • It's important to note that the metabolism of FFAs can vary depending on physiological conditions, such as fasting or exercise, and metabolic disorders can disrupt this process.

Fig. 2 A schematic view of obesity-associated, fatty acid-induced insulin resistance and metabolic dysfunction in skeletal muscle.Fig. 2 A schematic view of obesity-associated, fatty acid-induced insulin resistance and metabolic dysfunction in skeletal muscle. (Tumova, Jana, Michal Andel, and Jan Trnka, 2016)

Free Fatty Acids in Blood

The concentration of FFAs in the bloodstream is influenced by multiple factors, including dietary intake, lipolysis rates, hormonal regulation, and metabolic disorders. Elevated levels of FFAs, known as hyperlipidemia, have been associated with insulin resistance, obesity, and cardiovascular diseases. On the other hand, low levels of FFAs can also have detrimental effects on health, such as impaired hormone synthesis and reduced cell membrane integrity.

Free Fatty Acid Functions

FFAs serve as an essential energy source for various tissues, especially during prolonged fasting or intense physical activity. Additionally, FFAs act as signaling molecules, binding to specific receptors and regulating gene expression, inflammation, and insulin sensitivity. Furthermore, FFAs play a crucial role in lipid metabolism, serving as building blocks for the synthesis of complex lipids, including phospholipids, triglycerides, and cholesterol esters.

Free Fatty Acid Measurement

Accurate measurement of FFAs is crucial for understanding their physiological and pathological roles. Several methods are employed for the quantification of FFAs, including colorimetric assays, high-performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (GC-MS), and nuclear magnetic resonance (NMR) spectroscopy. These techniques allow researchers and healthcare professionals to monitor changes in free fatty acid profiles and assess their association with various diseases. Here are the main steps.

  • Sample Preparation: Determine the type of sample (e.g., plasma, serum, food product) and select an appropriate extraction method to release FFAs. Common techniques include solvent extraction, solid-phase extraction, or protein precipitation.
  • Method Optimization: Optimize the chosen method to enhance sensitivity, selectivity, and accuracy. This may involve adjusting parameters such as sample volume, extraction solvent, column type, mobile phase composition, or detection wavelength.
  • Data Analysis: Perform data analysis using appropriate software or calculations specific to the chosen analytical technique. Quantify the concentration of FFAs in the sample based on the calibration curve.
  • Interpretation: Interpret the results in the context of your research or application. Compare the measured concentrations to relevant reference values or establish correlations with other parameters of interest.

FFAs play a multifaceted role in human physiology, acting as both a vital energy source and important signaling molecules. Understanding the structure, metabolism, presence in the bloodstream, functions, and measurement methods of FFAs is crucial for unraveling their impact on health and disease. Learn about the importance of accurate measurement of FFAs and the methods employed for their quantification. Discover how measuring FFAs can provide insights into various physiological and pathological processes. Trust Creative Proteomics, a leading research organization, for expertise in free fatty acid analysis. You can get more information through our free fatty acid analysis service.


  1. Calder, Philip C. "Functional roles of fatty acids and their effects on human health." Journal of parenteral and enteral nutrition 39 (2015): 18S-32S.
  2. Tumova, Jana, Michal Andel, and Jan Trnka. "Excess of free fatty acids as a cause of metabolic dysfunction in skeletal muscle." Physiological research 65.2 (2016): 193.
  3. Kimura, Ikuo, et al. "Free fatty acid receptors in health and disease." Physiological reviews (2019).

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