We investigated the effect of Arogh, a polyherbal formulation (PHF) on animal models of anxietybased on exploratory behavior. The anxiolytic activity of polyherbal formulation (30, 100, 300 and500 mg/kg) was studied using various behavioural paradigms such as elevated plus maze (EPM),light/dark apparatus (LDA), open field apparatus (OFA), hole board apparatus (HBA). Diazepam(1 mg/kg) was used as a standard anxiolytic drug. The effect of PHF (100 and 300 mg/kg) onserotonin, dopamine and noradrenaline mediated behaviour was studied by lithium inducedhead twitches in rats, haloperidol induced catalepsy in mice and clonidine induced hypothermiain rats respectively. In EPM, PHF (100, 300 and 500 mg/kg) significantly (P < 0.05) increased thetime spent in open arms and the number of entries in open arms. In LDA, PHF (100, 300 and 500 mg/kg)significantly (P < 0.05) increased the time spent in lit zone. In OFA, PHF (100, 300 and 500 mg/kg)significantly (P < 0.05) increased the number of assisted rearing and the number of squarestraversed. In HBA, PHF (100, 300 and 500 mg/kg) significantly (P < 0.05) increased the number ofhead poking. In lithium induced head twitches, PHF (100 and 300 mg/kg) significantly (P < 0.05)decreased the number of head twitches. In haloperidol induced catalepsy, PHF (300 mg/kg)decreased the duration of catalepsy significantly (P < 0.05) at 60 min. In clonidine-inducedhypothermia, PHF (300 mg/kg) did not modify the effect. Drugs must be carefully assessed onEPM test and therefore in the present study EPM is supported by other tests. Present studyindicates that Arogh, a polyherbal formulation possess anxiolytic activity. It diminished serotonergictransmission and decreased the duration of catalepsy indicating potentiation of dopaminergictransmission. Thus, Arogh a polyherbal formulation contains bioactive principles which possessanxiolytic activity and modified 5-HT and DA mediated behaviour.