Plasma lipid levels and lipoprotein lipase (LPL) are known to follow circadian rhythms in rats. However, very little information is available on the variations in respiratory quotient (RQ) during the 24-h period in rats. The aims of this study were to provide an overall view of the effects of circadian rhythm on RQ and to determine the relationship of LPL and RQ with metabolic parameters in these animals. Male rats were fed ad libitum and were kept under a 12 : 12-h light–dark cycle. Rats were killed every 2 h over a 24-h period for measurement of metabolic parameters and tissue LPL activity. The RQ was measured every 4 h over the same 24-h period. The gastric contents increased during the dark phase and decreased during the light phase. For the metabolic parameters, circadian rhythms were detected for plasma glucose, triglycerides, high-density lipoprotein cholesterol and non esterified free fatty acids, but not for plasma total cholesterol or phospholipids. The RQ and adipose tissue LPL activity increased during the dark phase, while skeletal muscle LPL activity decreased during this phase. The RQ was inversely correlated with skeletal muscle LPL activity (r=−0.880) and positively correlated with adipose tissue LPL activity (r=0.937). These results appear to show that rats tend toward consumption of fat by accelerating fat oxidation, resulting in suppression of fat accumulation in the light phase, while tending toward fat accumulation by the suppression of fat oxidation in the dark phase.