Lippia origanoides H.B.K. (Verbenaceae) is a medicinal plant used in traditional medical practices for treatment of respiratory and gastrointestinal diseases. Based on previous reports regarding cardiovascu-lar effects induced by L. origanoides, this work aims to develop a standardized extract from L. origanoides aerial parts (Lo-HAE) and evaluate its hypotensive effect on mean arterial pressure in rats. Eight extraction systems were prepared by varying the ethanol/water ratio, sonication and time of extraction. The chromatographic profile and the determination of the flavonoid naringenin were performed by Ultra-fast Liquid Chromatography (UFLC) with UV detection at 290 nm. The extraction method for Lo-HAE was standardized considering the best extraction yield under 1:1 (v/v) ethanol/water ratio. Naringenin is the major compound of Lo-HAE, and then it was confirmed as the promising biomarker for Lo-HAE control assessment. For pharmacological studies, the acute oral toxicological assessment of Lo-HAE in female Wistar rats was performed. The Lo-HAE-induced hypotensive effect was evaluated by direct measure of pulse pressure after intravenous administration (12.5, 25 and 50 mg/kg), as well as indirect measure of blood pressure after oral administration (100 mg/kg and 200 mg/kg) in female Wistar rats. A marked decrease of mean arterial pressure was observed for Lo-HAE after both administration routes. In addition , no sign of either clinical or behavioral alterations was observed, as well as on the rats' body weight. The Lo-HAE demonstrates safe pharmacological potential for development of herbal medicines in the treatment of hypertension.