The root of Rotula aquatica is also called as pashanbed, belonging to the family Borogenaceae. It is widely distributed in India from kumaun to Assam and western to southern India.The medicinal values of plant lie in their component phytochemicals such as alkaloids, flavonoids, phenolic compounds and other nutrients like as amino acid, proteins, which produce a definite physiological action on the human body. The proximate values such as moisture content, ash value, extractive value, were carried out. The dried roots powder was subjected to successive extraction with different solvents like petroleum ether, benzene, chloroform, methanol, ethanol and water. Macroscopic as well as microscopic studies of any crude drug are the primary steps to establish its botanical quality control before going to other studies. Hence Standardization of crude drug plays a very important role in identifying the purity and quality of crude drugs. The present investigation reveals Standardization which include Moisture Content, Total ash, Acid insoluble ash, Water soluble ash, Water soluble Extractive value, Alcohol soluble Extractive value, Phytochemical screening, and further isolation and identification of Phytoconstituents.