Small sample sizes are typically incorporated in early Phase I clinical studies, which may lead to insignificant changes in safety parameters such as blood pressure. Therefore, it is paramount to identify an optimal, noninvasive method of accurately measuring blood pressure and an appropriate analysis strategy yielding the smallest variability. The goals of this study were (1) to compare the variability between automated and manual blood pressure measurements, (2) to determine whether triplicate blood pressure measurements were independent of one another, and (3) to assess how the number of blood pressure readings affects variability and study sample size. Twenty healthy volunteers were enrolled in this randomized, two-way crossover study. Each subject received three incremental infusions of phenylephrine or normal saline on separate days to simulate blood pressure variability. The mean systolic blood pressure readings with the automated device were consistently higher than the manual device by 3 to 5 mmHg. Conversely, the mean diastolic blood pressure readings with the automated device were consistently 3 to 5 mmHg lower than the manual device. However, the variability and absolute change in blood pressure were essentially identical with manual and automated methods. No systematic order effects such as the first blood pressure reading always being higher were detected, suggesting that the triplicate readings were independent of one another and that an interval of 2 minutes between readings is adequate. Compared to a single measurement, collecting blood pressure in triplicate results in a 40% lower sample size needed to detect a 5-mmHg difference in systolic blood pressure.