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Background: Rauvolfia serpentina mother tincture is the most widely used treatment for hypertension in homoeopathy. Rauvolfia serpentina contains the alkaloid Reserpine, which has been linked to depression. Objectives: To study acute oral toxicity and CNS depression of Rauvolfia serpentina MT (wild and cultivated) on wistar albino rats following OECD Guidelines 2001. Methods: Acute oral toxicity study was carried out for of Rauvolfia serpentina mother tincture (wild and cultivated). Overnight fasted healthy female wistar albino rats (n = 3) were administered orally Rauvolfia serpentina mother tincture (wild and cultivated) at a dose of 1 drop (Reserpine 0.25mg.), 10 drops (Reserpine 2.5 g.) and 30 drops (Reserpine 7.5 g.). After dosing, the animals were observed individually for the signs of toxicity and attention was directed to observations of tremors, convulsions, salivation, diarrhea, lethargy, sleep and coma. For modified Irwin test female wistar albino rats were divided into 5 groups of 8 animals each. The calculated dose of 2 drops in animal considering 20 drops in humans for 14 days (Reserpine 0.50 mg. /day) and calculated dose of 4 drops in animals considering 40 drops in humans for 14 days ( reserpine 1 gm. /day) was considered to rule out CNS depression in wistar albino rats. Results: In acute oral toxicity there was a significant decrease in motor activity and lost the righting reflex when Rauvolfia serpentina (wild and cultivated) is administered to wistar albino rats at a dose of 30 drops (reserpine 7.5 g.) and there was also mortality observed in all the 3 animals of this group when compared with other groups. In modified Irwin test there was CNS depression observed after 7 days when Rauvolfia serpentina (wild and cultivated) was administered to wistar albino rats at a dose of 4 drops per day for 14 days (Reserpine 1 gm. /day for 14 days). Nevertheless, Rauvolfia serpentina (wild and cultivated) at a dose of 2 drops per day for 14 days (Reserpine 50 mg. /day for 14 days) did not cause CNS depression. Conclusions: The acute oral toxicity was observed after administration of Rauvolfia serpentina MT (wild and cultivated) at dose of 30 drops (reserpine 7.5 g.). The further study confirms that the CNS depression begun to observed in wistar albino rats on the seventh day onward following the administration of Rauvolfia serpentina MT at a dose of 4 drops per day for 14 days (Reserpine at the dose of 1 gm. /day for 14 days). Consequently authors can conclude that dosage of Reserpine is an important factor in development of depression. Hence the preclinical safety pharmacology studies for neurobehavioral assessment are essential in order to selection of dose and safeguard patients against the negative effects of the medications used in clinical trials.