To define the contrast enhancement phases in the liver with perflubutane microbubbles, the liver enhancement time-intensity curves were investigated in 14 healthy volunteers. The agent was injected intravenously as a bolus and the liver was imaged with an ultrasound scanner as long as 4h after the injection. Time-intensity curves from the hepatic artery, the intrahepatic portal vein, the hepatic vein and the parenchyma of the liver were obtained from the liver ultrasound images. The arrival of the agent in the hepatic artery, the portal vein and the hepatic vein were visually distinguishable and the mean arrival times were 19.2, 24.3 and 32.2 s after the injection, respectively. The signal intensity in these vessels increased rapidly after the arrival of the contrast and gradually reverted to baseline after the peak. In contrast, within 5 min after the injection, the intensity in the parenchyma increased and reached a plateau, which persisted for at least 2h. The contrast enhancement phases in the liver with perflubutane microbubbles could be defined as two major phases-a vascular phase, in which the vessels are enhanced between 15 s and 10 min after injection, and a Kupffer phase, in which the parenchyma is enhanced 10 min after injection. The vascular phase is divided into three subphases: the arterial phase (15 to 45 s after injection); the portal phase (45 s to 1 min after injection); and the vasculo-Kupffer phase (1 to 10 min after injection).