Objective To investigate the effect of optimizing calculation method of insulin dosage in hypoglycemia growth hormone stimulation test [insulin tolerance test (ITT)] in patients with hypopituitarism.Methods Totally 106 patients with hypopituitarism admitted from February 2021 to March 2023 were selected for ITT, including 79 males and 27 females aged 14 to 61 years. Insulin dosage obtained by insulin optimization formula was used in the test, which is, insulin dosage (U) = body weight ×{-0.034+0.000 176×[0.5×(fasting insulin +180 min insulin)+60 min insulin +120 min insulin]+0.009 846×body mass index (BMI)}. The blood glucose changes of patients were monitored 30 min before the test, immediately after intravenous infusion and 30, 45, 60, 90 and 120 min after intravenous infusion, respectively, and the main complaints of patients were recorded.Results All 106 patients completed the test without any severe clinical reactions such as hypoglycemic coma or pituitary crisis. Among them, 76 patients (71.70%) experienced hypoglycemia within 30 min after the start of the test, 22 patients (20.75%) experienced hypoglycemia at 45 min, and the remaining 8 patients (7.55%) did not achieve a successful stimulation response. Comparison of the area under the glucose curve (AUC_Glu) among the 106 patients revealed that patients with simultaneous deficiencies in two hormones (growth hormone and cortisol) had lower overall blood glucose levels than those with a single hormone (growth hormone) deficiency. All patients’ blood glucose levels returned to normal by the end of the test.Conclusions The use of an optimized insulin dosage calculation method in ITT results in an earlier onset of hypoglycemia. This underscores the importance of thorough pre-test assessment by nurses, close monitoring of patients’ subjective complaints and blood glucose levels during the test, and proper nursing care to ensure the safety and success of the procedure.