Rapid Analysis of 37 FAMEs with the Agilent 8860 Gas Chromatograph

Significance of the Topic

Fatty acids are key constituents of neutral fats, phospholipids, and glycolipids, playing essential roles in nutrition and health. Analysis of their methyl esters (FAMEs) aids in quality control of foods and nutritional products, providing insights into saturated, monounsaturated, and polyunsaturated fatty acids.

Aims and Overview of the Study

This work demonstrates a rapid GC method using the Agilent 8860 system for simultaneous separation and quantification of 37 FAMEs in standard mixtures and infant proteolytic formulas, achieving faster runtimes than the GB 5009.168-2016 method with excellent resolution.

Methodology

• Sample derivatization by hydrolysis and methylation following national guidelines.
• Comparison of two DB-FastFAME capillary columns: 30 m×0.25 mm, 0.25 μm and 20 m×0.18 mm, 0.2 μm.
• Carrier gas: nitrogen at constant pressure; injection volume: 1 μL; split ratio 100:1.
• Oven temperature programs optimized for each column to maximize separation of cis/trans isomers and long-chain polyunsaturated fatty acids.

Used Instrumentation

• Agilent 8860 Gas Chromatograph with split/splitless inlet and flame ionization detector.
• Agilent 7693A automatic liquid sampler with a 5 μL syringe.
• DB-FastFAME capillary columns (30 m×0.25 mm, 0.25 μm, p/n G3903-63011; 20 m×0.18 mm, 0.2 μm, p/n G3903-63010).

Main Results and Discussion

• The 30 m column resolved all 37 FAMEs in under 24 minutes; the 20 m column completed separation in under 14 minutes, with resolution (Rs) greater than 1.8 for critical cis/trans pairs and omega-3 fatty acids (ARA, EPA, DHA).
• Calibration curves for ARA, EPA, and DHA displayed linearity (R² ≥ 0.9997) over 5–1000 mg/L concentration range.
• Method repeatability (n=6 injections) achieved area RSD below 1% for all analytes except C4:0.
• Analysis of amino acid, deep hydrolysis, and partial hydrolysis proteolytic formulas confirmed accurate detection and quantification of key unsaturated fatty acids in complex matrices.

Benefits and Practical Applications

The described method offers significantly reduced analysis time and maintains high chromatographic resolution, making it ideal for routine quality control in food, nutrition, and pharmaceutical laboratories, particularly for infant formula and fish oil analysis.

Future Trends and Opportunities

Advances in direct column heating, shorter high-efficiency columns, and fully automated sample handling are expected to further accelerate FAME profiling. Expansion into broader lipidomics applications and integration with high-throughput workflows will enhance analytical capabilities in research and industry.

Conclusion

The Agilent 8860 GC system paired with DB-FastFAME columns delivers a robust, rapid, and reproducible solution for comprehensive analysis of 37 fatty acid methyl esters. Choice of column length allows users to balance runtime, resolution, and column longevity according to analytical demands.

References

1. Zhou, Y.; Wu, H. Improving the analysis of 37 fatty acid methyl esters, Agilent Technologies Application Note, publication number 5991-8706EN, 2018.
2. Zhou, Y. Rapid separation of fatty acid methyl esters, Agilent Technologies Application Note, publication number 5994-0116EN, 2018.