A possibility to assess body composition during pregnancy is often important. Estimating body density (DB) and use the two-component model (2CM) to calculate total body fat (TBF) represents an option. However, this approach has been insufficiently evaluated during pregnancy. We evaluated the 2CM, and estimated fat-free mass (FFM) density and variability in 17 healthy women before pregnancy, in gestational weeks 14 and 32, and 2 weeks postpartum based on DB (underwater weighing), total body water (deuterium dilution) and body weight, assessed on these four occasions. TBF, calculated using the 2CM and published FFM density (TBF(2CM)), was compared to reference estimates obtained using the three-component model (TBF(3CM)). TBF(2CM) minus TBF(3CM) (mean ± 2SD) was -1.63 ± 5.67 (p = 0.031), -1.39 ± 7.75 (p = 0.16), -0.38 ± 4.44 (p = 0.49) and -1.39 ± 5.22 (p = 0.043) % before pregnancy, in gestational weeks 14 and 32 and 2 weeks postpartum, respectively. The effect of pregnancy on the variability of FFM density was larger in gestational week 14 than in gestational week 32. The 2CM, based on DB and published FFM density, assessed body composition as accurately in gestational week 32 as in non-pregnant adults. Corresponding values in gestational week 14 were slightly less accurate than those obtained before pregnancy.