The purpose of this investigation was to determine the accuracy of hand-to-hand bioelectrical impedance analysis (BIA) for estimating body composition in college-age female athletes using dual-energy X-ray absorptiometry (DEXA) as the criterion measure. Forty National Association for Intercollegiate Athletics college female athletes volunteered to participate in this study. For each participant, total body fat percentage (BF%) and fat-free mass (FFM) were obtained via BIA and DEXA. The mean BF% and FFM values obtained by BIA were compared with the criterion DEXA measure. The DEXA strongly correlated to the BIA for BF% (r = 0.74, R2 = 0.55, SEE = 3.60, and p < 0.01) and FFM (r = 0.84, R2 = 0.71, SEE = 2.45, p < 0.01). However, when compared with the DEXA, the mean values for BIA were significantly lower for BF% (DEXA = 27.6 ± 5.3%, BIA = 22.5 ± 3.5%, p < 0.01) and higher for FFM (DEXA = 47.2 ± 4.5 kg, BIA = 50.6 ± 4.6 kg, p < 0.01). The results of this investigation indicate that hand-to-hand BIA significantly underestimates BF% and overestimated FFM in college-age female athletes when compared with the criterion DEXA. Practitioners should use caution when analyzing body composition with hand-held BIA in a population of athletic women.