We analyzed two kinds of dairy fat differing in their contents of cis9, trans11-conjugated linoleic acid (cis9,trans 11-CLA or rumenic acid), and determined the positional distribution of this CLA-isomer within the three sn- positions of the triacylglycerol. In the high rumenic acid fat (HR), the CLA-isomers amounted to 2.1% of total fatty acids, and 0.8% in the low rumenic acid fat (LR). Over 90% of the total CLA-isomers were in the form of rumenic acid, with an identical isomeric CLA distribution in both fats. The two fats differed mainly with regards to their contents in palmitic acid, alpha-linolenic acid, and isomers of trans-C 18:1. Conversely, our stereospecific determination indicates that the positional distribution of rumenic acid is preserved among both types of fat, and is more specific for the sn-3 position of the triacylglycerol (54 to 64% of the total rumenic acid). Such a positional distribution is believed to be nutritionally relevant.