In previous work, we demonstrated that part of arachidonic acid (AA, n-6) pools in the gastrointestinal tract and blood forming tissues could be formed by local interconversion of linoleic acid taken up as free fatty acid from blood. In the rat the liver releases significant amounts of 2-arachidonyl-lysophosphatidylcholine (2-AA-LPC). In this study, we took advantage of the high hydrolytic activity of recombinant guinea pig pancreatic lipase-related protein 2 against the 1-ester bond of glycerophospholipids to conveniently prepare 2-[ 14C]AA-LPC by hydrolysis of radioactive phosphatidylcholine (L-α-1-Palmitoyl-2-arachidonyl [arachidonyl-1- 14C]) and examined the rate of uptake of 2-[ 14C]AA-LPC by the tissues in rats. Albumin-bound 2-[ 14C]AA-LPC was injected intravenously into male rats. The clearance rate of the 2-[ 14C]AA-LPC was fast, the initial half-life being 1.07 min. At 10 min after injection, the retention of 14C was 2.0–3.3%/g tissue of injected dose in the liver, 0.4 to 0.6%/g in stomach, 1.1 to 1.3%/g in small intestine, 0.4 to 0.6%/g in colon, 0.3 to 0.4%/g in bone marrow, and 0.7 to 1.0%/g in spleen. In most tissues, the proportion of radioactivity in tissue lipids was highest in PC (51 to 72%). In the heart, lung, and spleen the radioactivity in LPC was most predominant. The rate of uptake of AA as LPC by the liver was 61.0 ± 7.3 nmol/min, and by the gastrointestinal tract including stomach, small intestine and colon was 25.0 nmol/min. This study provides a quantitative estimation of the role of 2-AA-LPC in the supply of AA to the AA pools in extrahepatic tissues. It indicates that uptake of liver derived 2-AA-LPC may be one of the most important AA sources for extrahepatic tissues in the rat, particularly the gastrointestinal tract.