Method development and Identification of Triacylglycerols species with Supercritical Fluid Chromatography and Method Scouting Software

Significance of the Topic

Analysis of triacylglycerols (TAGs) is critical for assessing the quality and functionality of natural oils and fats. Conventional techniques can struggle to resolve the complexity of TAG species. Supercritical Fluid Chromatography (SFC) paired with mass spectrometry offers faster separation, reduced solvent usage and enhanced resolution for comprehensive TAG profiling.

Objectives and Study Overview

This study aimed to establish and optimize an SFC/MS/MS method for the identification and quantification of TAG species in edible oils. Method Scouting Software was employed to systematically explore key chromatographic parameters and accelerate method development.

Methodology and Instrumentation

The analytical workflow combined supercritical CO2 with acetonitrile modifier, a back pressure regulator and column oven to fine-tune retention and selectivity. MS/MS detection used ammonium adduct precursor ions in multiple reaction monitoring mode, targeting neutral losses of fatty acid moieties. Method Scouting Solutions software automatically generated 27 conditions by varying oven temperature, modifier concentration and system pressure to identify optimal settings.

Results and Discussion

Systematic scouting revealed the influence of each parameter on peak resolution and elution order. The optimized configuration (25 °C, 10 % acetonitrile, 10 MPa) provided baseline separation of palm oil TAGs, enabling accurate composition analysis. Applying this protocol to fully hydrogenated rapeseed oil shortened elution times for long-chain saturated TAGs, demonstrating method versatility.

Benefits and Practical Applications

• Rapid and high-resolution TAG profiling for quality control in food and oils industries
• Reduced solvent consumption compared with traditional LC methods
• Automated method optimization accelerates development time

Future Trends and Applications

Further integration of automated scouting with high-resolution mass analyzers and machine learning could expand lipidomics capabilities. Potential applications include trace-level detection of minor TAGs, authenticity testing and functional ingredient characterization.

Conclusion

The use of SFC/MS/MS combined with dedicated scouting software delivers an efficient and robust platform for detailed TAG analysis in natural oils, offering significant improvements in speed, resolution and solvent economy.

References

• Nakajima K, Zhe S, Mei hui J, Chee Keat Y, Yu Hua W, Nishimoto R. Method development and Identification of Triacylglycerols species with Supercritical Fluid Chromatography and Method Scouting Software. 2022 AOAC Annual Meeting.