The purpose of the present study was to investigate the effect of the combined administration of hepatic gluconeogenic substrates (glycerol + L-lactate + L-alanine + L-glutamine) on glucose recovery during insulin induced hypoglycemia (IIH), in rats. IIH was obtained by an ip injection of regular insulin (1 U/kg). Thus, 150 min after insulin administration the rats received an ip injection of glycerol + L-lactate + L-alanine + L-glutamine (each 100 mg/kg). In these experiments control groups, which received saline, glucose or isolated precursors (100 mg/kg), were employed. Glycemia was measured 30 min later, i.e., 180 min after insulin injection. The results showed that the combined administration of gluconeogenic precursors is more efficient than that of glucose itself to promote glycemia recovery. Since, the blood levels of hepatic glucose precursors were decreased (glycerol, L-lactate and L-alanine) or maintained (L-glutamine) during IIH, the ability of the liver to produce glucose from these gluconeogenic substrates was investigated. The results showed that the maximal capacity of the liver to produce glucose from glycerol (2 mM), L-lactate (2 mM), L-alanine (5 mM) and L-glutamine (5 mM) was increased. To L-alanine and L-glutamine, not only the glucose production was increased (P < 0.05) but also the production of L-lactate, pyruvate and urea. Therefore, the results suggest that the decreased availability of glucose precursors, promoted by insulin administration, limits the participation of hepatic gluconeogenesis to glycemia recovery. However, the administration of gluconeogenic precursors could overcome this limitation and promote better glycemia recovery than glucose itself.