Abstract: Background and Objective: Celastrus paniculatus-Willd belongs to the family Celastraceae is an endangered medicinal plant in India. It has potential role in primary health care system and various herbal drug formulations. The curative property of this medicinal plant is mainly due to the presence of various bioactive compounds. The present investigation was focused on the phytochemical screening of in vitro raised clones and mother plant in order to ensure the qualitative chemical similarity. Materials and Methods: The preliminary phytochemical screening such as fluorescence analysis of the leaf powder, fluorescence analysis of leaf extracts and various physiochemical properties of both in vitro raised clones and mother plant were done. To ensure the presence of various secondary metabolites TLC analysis was performed. The developed plates were sprayed with Dragendroff reagent for alkaloids, fast blue salt reagent for detection of phenolic group, ninhydrin for aminoacids and biogenic amines, vanillin phosphoric acid for detection of terpenoids. In order to confirm the qualitative chemical similarity of in vitro raised clones high performance liquid chromatography was performed. The spectral comparison of extracts was recorded by WIN CATS (1.3.4 version) software. The results were recorded and expressed as the Mean±SD for all the experiments. Results: The preliminary phytochemical screening of leaf extracts of both in vitro raised clones and mother plant exhibited the same result and there was no remarkable difference. TLC based identification of bioactive constituents revealed the presence of alkaloids, flavanoids, steroids, saponins, tannins, phenols, glycosides and reducing sugars. The HPTLC profile of the methanolic leaf extracts revealed the true to type nature of in vitro raised clones. Conclusion: The study revealed the presence of various bioactive constituents in both in vitro raised clones and mother plant of Celastrus paniculatus. The qualitative chemical comparison was done by TLC and HPTLC analysis.