Introduction. The assessment of body composition is relevant to establish nutritional status and identify potential health risks. Objective. a) To develop regression equations to predict fat mass (FM) using a dual-energy X-ray absorptiometry as reference method; b) to propose reference FM values based on chronological and biological age for Chilean children and adolescents. Methodology. Cross-sectional study in children and adolescents aged 5.0 to 18.9 years from the Maule Region (Chile). The sample was made up of 3593 subjects in a probabilistic fashion (stratified). Subjects' weight, standing height, sitting height, and waist circumference were assessed. Body mass index and age at peak development velocity (APGV) were estimated. Body composition (FM, fat-free mass, bone mass, and fat percentage) were established based on a dual-energy X-ray absorptiometry scan. Results. APGV (biological age) was 14.9 ± 0.9 years among boys and 11.5 ± 0.7 among girls. Equations were developed to estimate FM among boys and girls using chronological age, APGV, and waist circumference as predictors. Percentiles were estimated to assess FM by dual-energy X-ray absorptiometry and regression equations. Conclusion. Equations were acceptable to establish FM; in addition, reference values were proposed to assess FM based on chronological and biological age.