High-resolution triacylglycerol mixture analysis using high-temperature gas chromatography / mass spectrometry with a polarizable stationary phase, negative ion chemical ionization, and mass-resolved chromatography.

High-temperature gas chromatography/mass spectrometry (HT-GC/MS) has been employed to study the behavior of mixtures of triacylglycerol molecular species on a polarizable stationary phase (immobilized 65% phenyl methyl silicone). The use of negative ion chemical ionization (NICI) at an ion source block temperature of 300 °C overcomes problems with interpretation of electron ionization (EI) mass spectra produced during the HT-GC/MS. The NICI spectra of triacylglycerols produced under these conditions contain abundant [RCO2](-), [RCO2 - 18](-), and [RCO2 - 19](-) ions, believed to be produced by nucleophilic gas-phase ammonolysis, that are used to identify the individual fatty acid moieties associated with peaks in triacylglycerol total ion chromatograms. The polarizable stationary phase produces significantly enhanced resolution of triacylglycerol molecular species compared to hightemperature stable apolar stationary phases, such as immobilized dimethyl polysiloxanes. The resolution of complex natural mixtures of triacylglycerols can be further improved by use of the Biller-Biemann enhancement technique to produce mass-resolved chromatograms. Investigation of the mass-resolved chromatograms provides important information with regard to the factors that affect elution orders of individual triacylglycerol molecular species. The analysis of mixtures of authentic triacylglycerols by HT-GC/MS via NICI provides data that relate to the analytical limits of the technique for the analysis of triacylglycerols that bear both saturated and polyunsaturated fatty acyl moieties.