The study aims to identify the fatty acid composition in Prinsepia utilis seeds collected from diverse populations of Western Himalaya, India. The seeds contained higher saturated fatty acid (SFA-43.09%; Average of all populations) followed by polyunsaturated fatty acid (PUFA - 29.57%) and monounsaturated fatty acid (MUFA - 25.53%), however variations among growing populations was measured. The correlation analysis reveals a significant (p < 0.05) negative connection between 10-Octadecenoic acid methyl ester (FA1), and altitude. Edaphic factors like available nitrogen showed significantly (p < 0.05) but the negative correlation with cis-11-Eicosenoic acid, methyl ester (FA10; p < 0.001), Docosanoic acid, methyl ester (FA11; p < 0.05), Heptadecanoic acid, methyl ester (FA14; p < 0.05), and Tricosanoic acid, methyl ester (FA29; p < 0.05). Antioxidant activity did not show any variation in the seed oil of different populations but varied among the concentrations (5-50 µL) used. IC₅₀ values of DPPH activity was significantly (p < 0.05) varied connect ranged from 11.31-24.21 µL/mL among the different populations. These variations in fatty acid composition and IC₅₀ values of the DPPH activity can be one of the indicators for selecting a promising population that would be harnessed for its potential in nutraceuticals and cosmeceuticals industries.