Plants may favorably or adversely affect other plants through allelochemicals. The objective of this study was to examine the allelopathic effects of P. pinnata, A. indica and Z. jujuba on phytochemical profile of Tinospora cordifolia. In current study auxiliary buds of plant were cultured on Murasinge and Skoog medium, proved to be the good for shoot as well as root induction. To ensure the quality of plants grown by culture the juice of the plants collected from various supports from the same garden was analyzed by thin layer chromatography (TLC). A total number of 11 spots were observed. Out of the 11 spots three of Rf value 0.192 (Tinosporaside), 0.551 (berberine) and 0.717 were found to be universally present in the plants used in the study, which can be used as chemical markers of the plant. Densitometric profiles of the chromatograms showed variable amounts of constituents in the juice of plants supported by various trees. Tinosporaside contents were found to be significantly high in the plant supported by A. indica tree as comparative the plant supported by wall. Berberine contents were found to be almost same in all the plants, a bit higher levels were observed in the plant supported by P. pinnata. Tinosporaside and berberine showed to have antioxidant properties. However, overall antioxidant activity was observed to be high because of highest levels of tinosporaside in the plant supported by A. indica. The TLC profiles of explants grown with different concentration of aqueous extract of A. indica were found to be highly affected when observed after 8 days of culture. The significant difference was observed in the tinosporaside contents after 15 days growth of tissue cultured plants in presence of A. indica root extracts.