The existence of an intermediate pool of ubiquinone in intact mitochondria of rat heart was investigated. 2. The incorporation of [3H-methyl]S-adenosylmethionine into ubiquinone-9 was not influenced by the co-synthesis of the intermediate, 3-nonaprenyl-4-hydroxybenzoate. 3. In the intermediate-depleted mitochondria, the synthetic rate of the intermediate, 3-nonaprenyl-4-hydroxybenzoate was similar to that of ubiquinone. 4. The possible existence of 3-nonaprenyl-4-hydroxybenzoate as a metabolic pool under physiological condition is discussed.