Solanesol, a C45 isoprenoid is useful both as a medicine by itself and also as an essential intermediate used to synthesize a side chain component of coenzyme Q10 (Ubiquinone Q10),an important cardiac drug. Although solanesol is present naturally in some other plants, tobacco is the only known commercially important source of solanesol. It was isolated from tobacco for the first time in 1956. Typically, Solanesol is quantified by HPLC with C18 column and acetonitrile / methanol mobile phase with concentration gradients. In the new method, solenasol and its esters were extracted from tobacco with the help of Soxlet apparatus using hexane, followed by saponification. The saponified material is further analysed using HPLC equipped with C8 column and using constant ratio binary mobile phase consisting of acetonitrile and methanol. The limits of quantification (LOQ) and limits of detection (LOD) also have been optimized and recoveries of more than 90 % could be achieved.