Ruellia tuberosa L. (Acanthaceae) is an important medicinal plant, traditionally used to cure diseases. People make a tea with the roots to treat kidney stones and urinary tract infections. Hence the present study was carried to promote the synthesis of shoot production from different explants such as axillary and terminal meristems. For shoots, MS medium supplemented with different combination of hormones such as Kinetin and BAP were used and for roots NAA and IBA were used. A maximum shoot length was obtained in kinetin and BAP with 2 mg/l concentration and maximum root length was obtained in NAA and IBA with 1mg/l concentration. The aim of this study was to estimate and compare the concentration of bioactive compounds between wild and Invitro studies of R.tuberosa plants. A marked decrease in the total carbohydrate, flavonoids, quinones, and terpenoids were observed between Invitro propagated and wild type plants. This study clearly indicates that the environmental factors play a crucial role in the plant metabolic activities.