Separation of molecular species of triacylglycerols by high-performance liquid chromatography with a silver ion column.

Molecular species representative of the wide range of triacylglycerols occurring in nature, ranging from relatively saturated fats such as that from sheep adipose tissue through polyunsaturated seed oils, including sunflower and linseed oils, to an oligounsaturated fish oil, have been resolved by high-performance liquid chromatography in the silver ion mode. The stationary phase consisted of an ion-exchange medium, which was a silica gel matrix with bonded sulphonic acid moieties, loaded with silver ions. The mobile phase for the more saturated fractions was a gradient of acetone into 1,2-dichloroethane-dichloromethane, then acetonitrile was introduced to elute polyunsaturated fractions. A mass detector was employed to monitor separations. Fractions were collected via a stream-splitter for identification and quantification by gas chromatography as methyl esters. Excellent resolution was obtained on the silver ion column with no contamination of fractions with silver ions, while the column was stable and retained its activity in prolonged use.