The time course of action of regular and NPH insulins injected sc was studied in 15 insulin-treated diabetics over a 24-h period during which they received a constant infusion of glucose. The blood glucose began to decline in 1.2 +/- 0.1 h (range, 0.5--2) and reached its nadir in 5.7 +/- 0.3 h (range, 4--8) after the sc injection of regular insulin. The peak effect of regular insulin usually persisted for several hours, and the total duration of action was 16.2 +/- 1.1 h (range, 9--24). Both the time of peak effect and the total duration of action were considerably prolonged compared to data provided in standard textbooks. Free insulin increased to a peak in 2.7 +/- 0.3 h (range, 1--4) after regular insulin injection and then returned to baseline by 8.8 +/- 0.96 h. Subcutaneous injection of NPH insulin decreased the blood glucose by 2.4 +/- 0.5 h (range, 1--7), with a maximal effect at 11.0 +/- 1.4 h (range, 5--19). The total duration of effect on blood glucose was 25.1 +/- 0.7 h (range, 20--29). These values are similar to those in standard textbooks. Although the total insulin levels increased after the injection of NPH insulin, there was very little if any elevation in free insulin. Recognition of the prolonged effect of regular insulin is important in establishing an insulin treatment regime for diabetic patients.