The aims of this study were to describe normative values and seasonal variation of body composition in female cyclists comparing female road and track endurance cyclists, and to validate the use of anthropometry to monitor lean mass changes. Anthropometric profiles (seven site skinfolds) were measured over 16 years from 126 female cyclists. Lean mass index (LMI) was calculated as body weight × skinfolds(-x). The exponent (x) was calculated as the slope of the natural logarithm of body weight and skinfolds. Percentage changes in LMI were compared to lean mass changes measured using dual-energy X-ray absorptiometry (DXA) in a subset of 25 road cyclists. Compared to sub-elite and elite cyclists, world class cyclists were (mean [95% CI]) 1.18 kg [0.46, 1.90] and 0.60 kg [0.05, 1.15] lighter and had skinfolds that were 7.4 mm [3.8, 11.0] and 4.6 mm [1.8, 7.4] lower, respectively. Body weight (0.41 kg [0.04, 0.77]) and skinfolds (4.0 mm [2.1, 6.0]) were higher in the off-season compared to the early-season. World class female road cyclists had lower body weight (6.04 kg [2.73, 9.35]) and skinfolds (11.5 mm [1.1, 21.9]) than track endurance cyclists. LMI (mean exponent 0.15 [0.13, 0.18]) explained 87% of the variance in DXA lean mass. In conclusion, higher performing female cyclists were lighter and leaner than their less successful peers, road cyclists were lighter and leaner than track endurance cyclists, and weight and skinfolds were lowest early in the season. LMI appears to be a reasonably valid tool for monitoring lean mass changes.