Rauwolfia serpentina L. Benth Kurz, known as Sarpagandha, is mainly known for its chemical constituent’s reserpine, which was widely used as an antihypertensive drug and a powerful sedative. I has important medicinal values. During present study phytochemical analysis of root and leaves of plant. The evaluation of presence and absence of indole alkaloids were carried out by TLC and HPLC methods. The methods also focused on the quantitative and qualitative determination of indole alkaloids. This work represents a first report of antiproliferative activity of Rauwolfia serpentina. Plant leaves and roots were extracted by using solvent like ethanol and the quantity of crude extracts obtained was 12.05%. The evaluation of indole alkaloids were done by using the methods like TLC and HPLC that indicated the presence of four different indole alkaloid derivatives like ajmalicine, ajmaline, yohimbine and reserpine in root extract of Rauwolfia serpentina. Further quantitative determination of Rauwolfia alkaloids was carried out by spectrophotometric analysis which resulted that Ajmalicine content was greater in leaf extract whereas reserpine, ajmaline and yohimbine were greater in root extract of plant.