Hypertension is one of the most common chronic diseases. Inhibitory activity of angiotensin I-converting enzyme (ACE) is effective on giving hypotensive effect. Jamfruit leaf (Muntingia calabura L.) was reported to have excellent hypotensive effect. This research was aimed to test in manner of in vitro the inhibitory activity of ACE using ACE kit-WST, total phenolic content, and total flavonoid from jamfruit leaf ethanol extract, hexane, ethyl acetate, and butanol fraction. The result showed that Jamfruit leaf extract had ACE inhibitory activity and the most active fraction was ethyl acetate fraction. IC₅₀ value of the most active fraction, ethyl acetate fraction was 0.63 µg/mL . Ethyl acetate fraction also provides most flavonoid and phenolic content with value of 10.91 mg/g extract QE and 74.90 mg/g extract GAE. Ethyl acetate fraction of jamfruit leaf had most flavonoid, phenolic compound, and ACE inhibitory activity.