The triacylglycerols of Crepis alpina oil were characterized because this oil has a high concentration of crepenynic (cis-9-odadecen-12-ynoic) acid, a fatty acid useful in the chemical synthesis of deuterated fats for human metabolism studies. The triacylglycerols were separated from the crude oil by solid-phase extraction. Resolution, quantitation and isolation of the individual triacylglycerols were performed by silver ion high-performance liquid chromatography on a commercial column, an acetonitrile in hexane isocratic mobile phase and flame-ionization detection. Isolated triacylglycerols were identified by capillary gas chromatography of their fatty acid methyl esters. Of the eleven eluted triacylglycerols of Crepis alpina oil, 85% included 35% tricrepenynoyl, 34% linoleoyldicrepenynoyl and 16% dilinoleoylcrepenynoyl glycerols. Triacylglycerols eluted according to the numbers of alkene and alkyne bonds. Elution times, resolution and quantitation were reproducible over a three-month period. The flame-ionization detector response required no response factors for quantitation of the triacylglycerols present in Crepis alpina oil. The silver ion chromatography system permitted the identification of 95% of the triacylglycerols compared to 70% of the triacylglycerols previously identified with reversed-phase high-performance liquid chromatography.