The triacylglycerol stereospecific structure was determined for the major plant oils containing ψ-linolenic acid (GLA): evening primrose oil (EPO), black currant oil (BCO), borage oil (BO), andMucor javanicus fungal oil (MJO). It was found that GLA, although not α-linolenic acid, resisted pancreatic lipase hydrolysis. Therefore, the 2-position analysis was determined using phospholipase C-generated 1,2-diacylglycerol and phospholipase A2-generated lysophosphatidylcholine. GLA was found to be concentrated in the 3-position of EPO and BCO, the 2-position of BO, and the 2- and 3-positions of MJO. In BCO, octadecatetraenoic acid (n-3), also a †-6 fatty acid, was distributed similarly to GLA, but α-linolenic acid was found predominantly in the 1-position. Linoleic acid was nearly evenly distributed in all positions of EPO and BCO but was concentrated in the 1-position of BO and the 2-position of MJO. Both palmitic and stearic acids were found predominantly in the 1-position of all of the oils. The results demonstrate similarities and differences in the positional distribution of fatty acids in GLA-containing oils.