is a significant therapeutic plant has a place with family apocynaceae contains in excess of 70 distinct sorts of chemotherapeutic agents and alkaloids which help in treating different illnesses. For the most part, it is known as Vincarosea, Ammocallisrosea and Lochnerarosea. There are numerous or more than 400 alkaloids present in plant, which are used as flavor, agrochemicals, pharmaceuticals, fragrance, ingredients, food addictives, and pesticides. To develop a validated high performance thin layer chromatographic method for the analysis of leaves and flower extracts of Sample solutions were applied onto the plates with automatic TLC sampler Linomat V (Camag, Muttenz, Switzerland) and were controlled by WinCATS software. Plates were developed in 10 x 10cm twin trough glass chamber (Camag, Muttenz, Switzerland). A CAMAG TLC scanner was used for scanning the TLC plates. Pre-coated silica gel aluminium plates 60F254. For vincristine, simultaneous estimation of vincristine was performed by HPTLC on a silica gel plate using toluene-methanol-diethylamine (8.75: 0.75: 0.5, v/v/v) as the mobile phase. The method was validated as per the ICH guidelines. The Rf value was found to be 0.76 for flower and 0.80 for leaves at 250 nm which shows the presence of vincristin in . In this research paper, a validated HPTLC Method has been developed for the analysis of leaves and flower extracts