Traditionally Laetiporus sulphureus, commonly known as "Chicken-of-the-Woods", is considered a delicacy in various parts of the world and recently has shown medical potential in several treatments such as antitumor, antimicrobial, anti-inflammatory, antioxidant, immune-modulating etc. However, quality standards of this species have not been studied so far. For this purpose, microscopic features of powder such as physical characters, spore measurement, type of hyphae were examined and recorded. Physicochemical parameters like organoleptic features, loss on drying (5.15 %), total ash (3.04 %), swelling capacity (1.66 ml/g), water holding capacity (163.16 g/g), water solubility index (21.73 %) and fluorescent behaviour against 16 reagents were also determined. In addition, alcoholic extract was prepared using methanol as solvent where extractive value was 24.27 %. Preliminary mycochemical analysis indicated presence of cardiac glycoside, flavonoid, phenol, saponin and terpenoids in the extract. While quantitatively major bioactive components were present in the following order of flavonoid > phenol > ascorbic acid > beta-carotene > lycopene. Moreover, HPTLC and HPLC profiles were recorded to determine phenolic fingerprint. HPTLC characterization showed presence of at least 5 components as detected by scanning the plate at 300 nm and HPLC chromatogram indicated existence of 32 peaks which might be of phenolic compounds. In addition, superoxide radical scavenging (EC50 1.38 mg/ml), DPPH radical scavenging (EC50 0.11 mg/ml), chelating ability of ferrous ion (EC50 0.27 mg/ml), reducing power (EC50 0.26 mg/ml) and total antioxidant capacity (18.01 A mu g AAE/mg of extract) were determined to evaluate antioxidant potentiality of the extract. The above parameters are significant towards establishing pharmacognostic standards for future authentication of genuine mushroom material.