FAME Analysis in Olive Oil and Seed Oils

Application Note:

Learn how fatty acid methyl ester profiling is used to assess olive oil and seen oil composition. Includes derivatisation steps, GC conditions, and comparative results for olive and sunflower oil.
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Introduction

Fatty acid methyl ester analysis is widely used to assess the composition and authenticity of edible oils. Olive oil is a high-value product, which makes it vulnerable to adulteration with cheaper seed oils such as sunflower or hazelnut.

FAME profiling measures the relative ratios of individual fatty acids within an oil sample. These ratios help confirm authenticity and provide nutritional information for labelling.

This method uses the Ellutia 200 Series Gas Chromatograph with FID detection and an ELR-CN100 column to separate and quantify fatty acid methyl esters in olive and seed oils

 

FAME_Analysis

The Challenge

 

Oil authenticity depends on accurate fatty acid ratios

 

Olive and sunflower oils differ primarily in the ratios of oleic acid and linoleic acid. However, both contain a range of saturated and unsaturated fatty acids, from palmitic acid to behenic acid

Adulteration does not always result in obvious visual differences. The composition must be measured analytically to identify changes in fatty acid distribution. Small shifts in ratio can indicate substitution or contamination.

To confirm authenticity, laboratories need a method that separates individual fatty acids clearly and produces consistent peak areas for accurate ratio calculation.

 

FAME_Profiling_1

The Solution

 

FID detection provides clear quantification of FAME profiles 


Gas chromatography separates fatty acid methyl esters based on chain length and degree of saturation. The flame ionisation detector provides stable and proportional response across the fatty acid range.

 

 

Ellutia_200_GC

 

In this study, oils were converted to their FAME equivalents prior to analysis. The Ellutia 200 Series GC, combined with the ELR-CN100 column, achieved separation across the full fatty acid profile.

Clear peak separation allows relative ratios to be calculated reliably, which helps identify differences between olive and sunflower oil.

 

Method Overview

 

How olive oil and seed oils are prepared and analysed

 

Sample Preparation
  • Weigh 1-25mg of oil into a reaction vessel 
  • Add 1mL toluene
  • Add 2 mL 10 percent sulfuric acid in methanol and shake.
  • Seal and heat for 30 minutes at 60 °C.
  • Cool, then add saturated sodium bicarbonate and hexane.
  • Dry the organic layer with anhydrous sodium sulfate.
  • Inject 1 µL into the GC
 
GC Conditions
  • Injector: 250 °C
  • Carrier gas: Hydrogen
  • Column flow: 1.0 mL/min
  • Split flow: 100 mL/min
  • Injection volume: 1.0 µL
  • Column: ELR-CN100 60 m × 0.25 mm × 0.2 µm
  • Initial temperature: 140 °C hold 5 minutes
  • Detector: FID at 250 °C

These conditions allow clear separation of palmitic, stearic, oleic, and linoleic acid methyl esters.

Results and Reliability

 

What the analysis shows


Chromatograms showed clear separation of the major fatty acids in olive and sunflower oil. Olive oil showed high oleic acid content and lower linoleic acid levels. Sunflower oil showed the opposite pattern, with higher linoleic acid and lower oleic acid.

The relative ratios calculated from peak areas demonstrated clear compositional differences between the two oils. These ratio differences provide a measurable way to assess authenticity and detect substitution.

Consistent peak areas and stable detector response allow repeatable profiling across multiple samples.

 

Learn More

 

Get the full method and results


If you’d like to see the full details behind this testing method, you can download the complete application note. It includes chromatograms, calibration data, and the exact conditions used for the analysis. It’s a handy reference if you want to check your own setup or compare results.

 

 

Download the full application note.

 

 

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FAME_Analysis_Olive_Seed_Oils-1
 

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