Propofol is frequently used for sedation, induction, and maintenance of anesthesia. It is, however, associated with pain on injection. Propofol-Lipuro® has an oil phase that allows a larger proportion of propofol to be dissolved in it and, thereby, apparently reduces pain. However, studies investigating this have had methodological limitations. We devised a randomized, double-blind, crossover study comparing pain on injection between two preparations of propofol, Diprivan® and Propofol-Lipuro®, in subanesthetic doses. Sixty healthy patients received the drugs in random order via the same injection site separated by 10 min and a 0.9% saline flush. Pain was assessed using a verbal rating score (VRS) during and at 1-min time points after injection. Differences in VRS between the two propofol preparations at different time points in each patient were analyzed. In patients who were given Diprivan® first followed by Propofol-Lipuro® (group D-P), pain was significantly reduced with Propofol-Lipuro® compared with Diprivan® during initial injection (median difference in VRS = 2 [interquartile range 0–2], P = 0.002) and at 1 min (3 [0–4], P < 0.001). In patients who were given Propofol-Lipuro® first followed by Diprivan® (group P-D), no significant differences in VRS were shown. Propofol-Lipuro® is associated with reduced injection pain compared with Diprivan® and also seems to attenuate subsequent injection pain of Diprivan® when administered first. The mechanism is unknown, but may be related to a reduction in the concentration of propofol in the aqueous phase.