Age-related loss of skeletal muscle mass results in a reduction in metabolically active tissue and has been related to the onset of obesity and sarcopenia. Although the causes of muscle loss are poorly understood, dietary fat has been postulated to have a role in determining protein turnover through an influence on both inflammation and insulin resistance. This study was designed to investigate the cross-sectional relation between dietary fat intake, as dietary percentage of fat energy (PFE) and fatty acid profile, with indices of skeletal muscle mass in the population setting. Body composition [fat-free mass (FFM; in kg)] and the fat-free mass index (FFMI; kg FFM/m(2)) was measured by using dual-energy X-ray absorptiometry in 2689 women aged 18-79 y from the TwinsUK Study and calculated according to quintile of dietary fat (by food-frequency questionnaire) after multivariate adjustment. Positive associations were found between the polyunsaturated-to-saturated fatty acid (SFA) ratio and indices of FFM, and inverse associations were found with PFE, SFAs, monounsaturated fatty acids (MUFAs), and trans fatty acids (TFAs) (all as % of energy). Extreme quintile dietary differences for PFE were -0.6 kg for FFM and -0.28 kg/m(2) for FFMI; for SFAs, MUFAs, and TFAs, these were -0.5 to -0.8 kg for FFM and -0.26 to -0.38 kg/m(2) for FFMI. These associations were of a similar magnitude to the expected decline in muscle mass that occurs over 10 y. To our knowledge, this is the first population-based study to demonstrate an association between a comprehensive range of dietary fat intake and FFM. These findings indicate that a dietary fat profile already associated with cardiovascular disease protection may also be beneficial for conservation of skeletal muscle mass.