Description:Diurnal oscillation in glucose metabolism is strictly regulated by various factors, including activity, sleep and diet. We used a isotopologue tracing method to characterize the diurnal oscillation in glucose metabolite isotopes in Drosophila sp. Wild type flies were injected with uniformly labeled 13C glucose. Different isotopologues of glucose metabolites were measured using UPLC-MS/MS to assess their temporal pattern. In addition, the effect of hyperactivity and sleep on the temporal variation of metabolite isotopologues was assessed using the hyperactive Fumin mutant flies.