Rauvolfia belongs to the family Apocynaceae. Rauvolfia tetraphylla is economically important medicinal plant because of the presence of various alkaloids. Alkaloid yohimbine is hypotensive a cardiovascular depressant and hypnotic. In our study we have taken the methanol extract of leaves, stem and roots of Rauvolfia tetraphylla and yohimbine as standard. The compression was done by HPTLC. The quantification and identification of yohimbine was done by WINCATS software with densitometric detection (Camag Scanner -3). The present technique allowed different separation profiles that can be useful for phytochemicals characterization of various parts of plants. The linear calibration ranges were 10-1000 g/ml for yohimbine. Yohimbine was detected only in the leaves of plant in 6.11 %. The HPTLC methods were successfully validated and applied to the quantization of yohimbine.