The purpose of the present study is to investigate the pharmacognostical characteristics of the roots of Saccharum officinarum Linn. (poaceae) to establish standardized parameters. Macroscopic, microscopic, and physicochemical evaluation; preliminary phytochemical analyses and high performance thin layer chromatography (HPTLC) profile were done for qualitative phytochemical evaluation. Roots are grey to blackish brown, fibrous, solid, jointed, odour and taste not distinct. Transverse section of both thin and fairly thick root showed presence of epidermal, hypodermal layers, aerenchymatous cortex and a wide stele. Powder microscopy revealed presence of long thick walled fibers with pointed ends, vessels and tracheids with reticulate, scalariform and bordered pitted walls and thin walled parenchyma cells. Physicochemical parameters viz. percentage of moisture content (9.3% w/w), total ash (7.5% w/w), acid insoluble ash (5.0% w/w), water soluble extractive (6.9% w/w) and ethanol soluble extractive (3.8% w/w) were also determined. Various groups of phytoconstituents found in the plant are carbohydrate, steroids, flavonoids, saponins, tannins and phenolic compound. HPTLC profile of hydroalcoholic extract showed presence of twelve different components in the solvent system, chloroform: methanol (9:1, v/v). The different salient diagnostic features established in this study will help for proper identification and standardization of the drug in crude form, and then the plant material can be used for further studies.