This study aimed to produce gluten-free cupcakes (GFCs) using a composite flour blend of Lepidium sativum seed mucilage (LSM) in different proportions. The GFCs substituted oil with 100% LSM (M100), revealing significant reduction in fat content (87.55%), protein 13.43 g/100 g, fiber 2.48 g/100 g, and carbohydrates 58.39 g/100 g. The M100 GFC showed favorable fatty acid ratios (1.80 MUFA:SFA, 0.95 PUFA:SFA, 0.97 omega 6:omega 9) and lipid health quality indices (0.72 atherogenicity index, 0.59 thrombogenic index, 1.39 health-promoting index, and 2.3% ∆desaturase index), with sensory acceptability at 7.07 ± 0.21. The LSM effectively replaced fat, enhanced functionality and improved sensory profiles 7.07 ± 0.21, suggesting its potential for industrial applications.