The objective of this study is to obtain reduced fat biscuits by using chia flour instead of vegetable fat and to investigate the effects of chia flour on the physicochemical, nutritional and sensory properties of biscuits. Fat was replaced by chia flour at the ratios of 25%, 50%, 75% and 100%. Fat replacement caused a significant decrease in fat, while significant increases in ash, protein, carbohydrate, total dietary fibre, phosphorus, magnesium, calcium, potassium, phenolic compounds, essential fatty acids (linoleic acid and linolenic acid) and hardness of biscuits were observed. Sensory quality was decreased with the increasing level of chia flour in biscuits. Biscuit containing 25% fat replacer was acceptable in terms of overall acceptance. This sample has 14.12% protein, 17.403% linoleic acid and 9.252% linolenic acid, while control has 10.51% protein, 13.068% linoleic acid and 7.751% linolenic acid. In addition, higher total dietary fibre, minerals and phenolic compounds were obtained. This study suggests that chia flour could be used in biscuit production to reduce fat and increase bioactive components of biscuits.