► The major constituents of R. officinalis fractions and essential oil were determined. ► Fractions and essential oil of R. officinalis reduced immobility time in the TST. ► Carnosol and betulinic acid produced antidepressant-like effect in the TST. ► Results support the notion that R. officinalis has antidepressant properties. The aim of this study was to investigate the antidepressant-like effect of fractions from Rosmarinus officinalis L.: ethyl acetate 1 and 2 (AcOEt1 and 2), hexane (HEX), ethanolic (ET), and essential oil-free (EOF) fractions, as well as essential oil, the isolated compounds carnosol and betulinic acid in the tail suspension test, a predictive test of antidepressant activity. Swiss mice were acutely administered by oral route (p.o.) with fractions, essential oil or isolated compounds, 60min before the tail suspension test or open-field test. All of them produced a significant antidepressant-like effect: AcOEt1, ET, EOF fractions and essential oil (0.1–100mg/kg, p.o); HEX (0.1–10mg/kg, p.o) and AcOEt2 fraction (0.1–1mg/kg, p.o), carnosol (0.01–0.1mg/kg, p.o.) isolated from the HEX fraction and betulinic acid (10mg/kg, p.o.), isolated from the AcOEt1 and AcOEt2 fractions. No psychostimulant effect was shown in the open-field test, indicating that the effects in the tail suspension test are specific. This study suggests that carnosol and betulinic acid could be responsible for the anti-immobility effect of extracts from R. officinalis.