Low water intake and underhydration are public health issues that may increase risk for diseases such as Type 2 Diabetes Mellitus. Studies suggest that high vasopressin (AVP) levels associated with low water intake may contribute to hyperglycemia. This study explored the physiological system by which AVP impairs glucose regulation through a single-blind randomized, counterbalanced, crossover design. This is a pilot and feasibility study of AVP infusion at increasing incremental rates, which was completed to determine the rate of infusion for the cross-over study. Participants completed a control and experimental trial. The experimental trial included a 3-hour AVP infusion and a 2-hour euglycemic-hyper insulinemic clamp at the end of the first hour versus control of 0.9% sodium chloride replacing AVP. In both trials, blood samples were taken every 5 minutes to measure glucose, as well as 7 other time points of insulin infusion. Two participants completed the pilot (47.5±3.5 years, 172.5 ±7.5cm, 82.5±17.7kg, 27.5±3.5 kg/m2, 5.1±0.64% HbA1c), and 3 participants completed the cross-over study (49±1.7 years, 173.7±6.7cm, 80.4±150kg, 26.5±3.2kg/m2, 5.3±0.2% HbA1c), all females. The rate of AVP infusion for the cross-over study was 12.5 mU/min. Compared to the control, the AVP trial blood glucose trended higher towards the end of the experiment, as did glucose metabolism, plasma osmolality, and plasma volume. Blood pressure was slightly higher in the AVP trial versus the saline, while plasma sodium and potassium levels did not differ. Total plasma protein seemed higher in the saline trials than in the AVP trials. This study supports the notion that increased levels of vasopressin over time may increase blood glucose. This could lead to supplementation of type 2 diabetes interventions with increased water intake.