We made a comparative analysis of the uptake, tissue deposition and conversion of dietary alpha-linolenic acid (ALA) to its long chain metabolites eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) with preformed EPA + DHA. Diets containing linseed oil [with ALA at approximately 2.5 (4 g/kg diet), 5 (8 g/kg diet), 10 (16 g/kg diet), 25% (40 g/kg diet)] or fish oil [with EPA + DHA at approximately 1 (1.65 g/kg diet), 2.5 (4.12 g/kg diet), 5% (8.25 g/kg diet)] or groundnut oil without n-3 polyunsaturated fatty acids (n-3 PUFA) were fed to rats for 60 days. ALA and EPA + DHA in serum, liver, heart and brain increased with increments in the dietary ALA level. When preformed EPA + DHA were fed, the tissue EPA + DHA increased significantly compared to those given ALA. Normalized values from dietary n-3 PUFA to tissue EPA + DHA indicated that 100 mg of dietary ALA lead to accumulation of EPA + DHA at 2.04, 0.70, 1.91 and 1.64% of total fatty acids respectively in liver, heart, brain and serum. Similarly 100 mg of preformed dietary EPA + DHA resulted in 25.4, 23.8, 15.9 and 14.9% of total fatty acids in liver, heart, brain and serum respectively. To maintain a given level of EPA + DHA, the dietary ALA required is 12.5, 33.5, 8.3 and 9.1 times higher than the dietary EPA + DHA for liver, heart, brain and serum respectively. Hence the efficacy of precursor ALA is lower compared to preformed EPA + DHA in elevating serum and tissue long chain n-3 PUFA levels.