Estimates of body fat mass gained during human pregnancy are necessary to assess the composition of gestational weight gained and in studying energy requirements of reproduction. However, commonly used methods of measuring body composition are not valid during pregnancy. We used measurements of total body water (TBW), body density, and bone mineral content (BMC) to apply a four-component model to measure body fat gained in nine pregnant women. Measurements were made longitudinally from before conception; at 8-10, 24-26, and 34-36 wk gestation; and at 4-6 wk postpartum. TBW was measured by deuterium dilution, body density by hydrodensitometry, and BMC by dual-energy X-ray absorptiometry. Body protein was estimated by subtracting TBW and BMC from fat-free mass. By 36 wk of gestation, body weight increased 11.2 +/- 4.4 kg, TBW increased 5.6 +/- 3.3 kg, fat-free mass increased 6.5 +/- 3.4 kg, and fat mass increased 4.1 +/- 3.5 kg. The estimated energy cost of fat mass gained averaged 44,608 kcal (95% confidence interval, -31, 552-120,768 kcal). The large variability in the composition of gestational weight gained among the women was not explained by prepregnancy body composition or by energy intake. This variability makes it impossible to derive a single value for the energy cost of fat deposition to use in estimating the energy requirement of pregnancy.