The ability of fetal pig skeletal muscle (biceps femoris) to metabolize glucose, fructose, lactate, acetate and palmitate in vitro was examined at 70, 90 and 110 d of gestation. Even though succinate dehydrogenase activity increased as fetal age increased (P less than .01), the rate of oxidation of glucose, fructose, acetate and palmitate to CO2 was not influenced by fetal age (P greater than .05), but each rate was dose-dependent (P less than .01). At higher concentration, lactate oxidation to CO2 proceeded at a faster rate in the muscle of 70-d fetuses when compared with 90- or 110-d fetuses. Muscle glycogen content increased (P less than .01) from 70 to 110 d of gestation. The rate of glucose incorporation into glycogen increased over this same time frame (P less than .06). Glucose-6-phosphate dehydrogenase activity did not change with age when activity was expressed per unit wet weight of muscle (P greater than .05). The ratio [1-14C]glucose/[6-14C]glucose oxidized to CO2 was independent of age and substrate concentration, and indicated significant pentose cycle activity in fetal muscle. Incorporation of lactate and palmitate into phospholipid was greatest at 70 d of gestation, a time period that coincides with establishment of mature muscle fiber number and fiber hypertrophy. The rate of palmitate incorporation into triacylglycerol was dependent on concentration of substrate (P less than .01) but not on age (P greater than .05). The rate of fructose oxidation to CO2 was lower than the rate for glucose, lactate and acetate when compared at similar concentrations. Acetate carbons were not incorporated into free fatty acids or lipid.(ABSTRACT TRUNCATED AT 250 WORDS)