The effect of alfalfa seed flour supplementation on the quality characteristics of refined wheat flour-based biscuits was studied. The proximate composition of refined wheat flour and alfalfa seed flour was determined. Refined wheat flour contained 12.43% moisture, 11.52% crude protein, 1.61% crude fat, 0.71% crude fiber, 1.43% ash and 70.83% nitrogen free extract, while alfalfa seed flour contained 5.79%, 29.49%, 12.71%, 5.53%, 4.80% and 41.73% moisture, crude protein, crude fat, crude fiber, ash and nitrogen free extract correspondingly. Alfalfa seed flour at 5%, 10%, 15% and 20% supplementation levels was incorporated in refined wheat flour to produce composite flour. The biscuits prepared were subjected to quality evaluation. Physical analysis of biscuits disclosed that supplementation of alfalfa seed flour decreased the width from 47.25 to 42 mm and the spread factor from 62.7 to 53.12, while it increased the thickness from 7.53 to 8.10 mm. Supplementation of refined wheat flour–based biscuits with alfalfa seed flour at different inclusion levels significantly (p < 0.05) increased the crude protein content from 10.19% to 15.30%, the crude fiber content from 0.73% to 1.62%, the crude fat content from 17.46% to 21.59% and the ash content from 1.37% to 1.92%, whereas it decreased the moisture content from 3.57% to 3.26% and the nitrogen free extract from 66.90% to 59.32%. The effect of supplementation on the mineral contents of biscuits was also significant (p < 0.05). Potassium, magnesium, calcium, iron and zinc contents increased from 105.30, 14.65, 43.91, 3.74 and 0.94 to 145.00, 26.64, 79.60, 7.93 and 1.60 mg/100 g, respectively. Sensory evaluation revealed that the quality score of biscuits in terms of color, taste, texture and overall acceptability decreased with increased supplementation. The present research work confirmed that a maximum of 10% alfalfa seed flour supplementation in refined wheat flour could produce acceptable biscuits with an appropriate nutritional profile.