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The present investigation was carried out to characterize four different types of psyllium seeds including one landrace for their nutritional, structural, antioxidant profile and biological active compounds. DSC analysis showed the highest peak temperature (TP) of 107.57 ºC in RI-89 and end set temperature (TE) of 130.2 ºC for HI-5 cultivar. FT-IR analysis displayed strong absorption bands at frequencies 960, 1380, 1740, 2280 and 3280cm-1. SEM micrographs of each cultivar showed compact and irregular mass of fiber at various magnifications. Protein, fat, ash and total carbohydrate content of psyllium seed flour were recorded up to 13.33, 0.38, 5.0 and 77.88 %, respectively. Ethanol was found to be a suitable solvent than methanol to extract the phytonutrients from psyllium seed flour. DPPH free radical scavenging activity (FRSA) of methanolic and ethanolic extract varied between 31 to 39 % and 38.9 to 43 %, respectively. However, ABTS-FRSA varied between 31.7 to 32.9 % and 33.2 to 34.3 %, for methanolic and ethanolic extract, respectively. Total flavonoids and total phenolic content was recorded up to 2.92 mg RE/g and 4.37 mg GAE/g, in ethanolic extract. The reducing power (RP) and Metal chelating activity (MCA) also varied significantly (p≤0.05) among the cultivars. The MCA ranged from 33.20 to 34.81 % in methanolic extracts and 34.62 to 36.07 % in ethanolic extracts. The lowest absorbance for reducing power was found in methanolic extract of landrace (0.16), whereas, the highest was found in ethanolic extract of RI-89 (0.37). Physico-chemical properties including oil and water absorption capacity of both seed and flour did not vary significantly among the cultivars, whereas significant difference was observed in seed hydration capacity among cultivars evaluated. Among the physical properties, L/W ratio ranged from 1.80 to 199, whereas porosity and angle of repose were of 59.2 to 67.7 % and 23.20 to 27.02º, respectively. Static coefficient of friction was found less using steel (0.484 to 0.667) as a test surface compared to wood (0.679 to 0.744) and plastic (0.536 to 0.560). Bulk density, true density and porosity of various psyllium seed cultivars varied between 0.57 to 0.63 g/cm3 and 1.50 to 1.77 g/cm3 and 58.2 to 67.7%, respectively. Geometric mean diameter (Dg), Arithmetic mean diameter (Da) and surface area of seeds (A) varied from 1.49 to 1.60 mm and 1.68 to 1.79 mm and 5.91 to 6.78 mm2, respectively.