Administration of exogenous insulin during an intravenous glucose tolerance test allows the use of the minimal model technique to determine the insulin sensitivity index in subjects with reduced endogenous insulin responses. To study the effect of different insulin administration protocols, we performed three intravenous glucose tolerance tests in each of seven obese subjects (age, 20-41 yr; body mass index, 30-43 kg/m2). Three different insulin administration protocols were used: a low-dose (0.025 U/kg) infusion given over 10 min, a low-dose (0.025 U/kg) bolus injection, and a high-dose (0.050 U/kg) bolus injection, resulting in peak insulin concentrations of 1,167 +/- 156, 3,014 +/- 483, and 6,596 +/- 547 pM, respectively. The mean insulin sensitivity index was 4.80 +/- 0.95 x 10(-5), 3.56 +/- 0.53 x 10(-5), and 2.42 +/- 0.40 x 10(-5) min-1/pM respectively (chi +/- SEM; P = 0.01). The association of higher peak insulin concentrations with lower measured insulin sensitivity values suggested the presence of a saturable process. Because results were not consistent with the known saturation characteristics of insulin action on tissue, a second saturable site involving the transport of insulin from plasma to interstitium was introduced, leading to a calculated Km of 807 +/- 165 pM for this site, a value near the 1/Kd of the insulin receptor. Thus, the kinetics of insulin action in humans in these studies is consistent with two saturable sites, and supports the hypothesis for transport of insulin to the interstitial space. Saturation may have an impact on minimal model results when high doses of exogenous insulin are given as a bolus, but can be minimized by infusing insulin at a low dose.