Background: Ethnomedicinally, the plant Argyreia pilosa Wight & Arn. (Convolvulaceae) has long been utilized in various disorders in the conventional system; most significantly it is utilized against sexually transmitted diseases, skin troubles, diabetes, rheumatism, cough, and quinsy. The key challenge experienced in the standardization of herbal drugs is the lack of proper identification of plant source. Therefore there is certainly have to establish quality control parameters by utilizing pharmacognostic and phytochemical evaluation, that ensure the purity, safety, and efficacy of medicinal plant A. pilosa. Aim: To assess pharmacognostic characteristics which include macroscopic, microscopic and physicochemical parameters of the root of A. pilosa. Methods: Micro and Macroscopic characters of fresh and dried root samples were investigated. Physicochemical parameters had been done by using WHO recommended parameters, preliminary phytochemical and fluorescent analysis of root sample were carried out for proper identification and standardization of root of A. pilosa. Results: The color, shape, size, odor, and surface characteristics were noted from the root and powdered root material of A. pilosa. Light electron microscope i.e., Olympus CX-21i trinocular Microscope images of cross section of root and powdered root revealed that the presence of cork cells, Xylem fibers with tapered ends, lignified xylem vessels, phloem fibers, medullary rays, sclerides and parenchymatous cells. Phytochemical screening showed the presence of flavonoids, alkaloids, tannins, phenols, steroids, acid compounds, glycosides, amino acids, and proteins. Physicochemical parameters such as moisture content, ash value, extractive value and fluorescent behavior of root powder were determined. These parameters are helpful to differentiate the powdered drug material. Conclusion: The current research is useful in order to supplement the information with regard to its standardization, identification and in carrying out further investigation in Ayurvedic system of medicine.