AbstractTriacylglycerols (TGs) of lipase‐modified butter oil were separated into saturated, monoene, diene and triene fractions on ap‐propylbenzene sulfonic acid solid‐phase extraction column loaded with silver ions. Fatty acid analysis of the fractions showed that the amounts of saturated TGs (98.4 mol%) and monoene TGs (26.0 mol%) in the saturated and monoene fractions, respectively, were close to the theoretical amounts of TGs in pure fractions. The column method provides a useful alternative to AgNO3‐thin‐layer chromatography as a means of separating the TGs of butterfat and producing relatively pure TG fractions for further analysis by gas chromatography (GC) or GC‐mass spectrometry.