Objective To examine changes in intra-tracheal fluid flow parameters during fetal breathing movements throughout the second half of pregnancy in the normally developing human fetus. Design Prospective cross-sectional study.Setting Fetal medicine unit at the Charité University Hospital in Berlin.Methods Assessment of tracheal fluid flow was attempted in 340 healthy fetuses (GA 20-40 weeks) in which fetal breathing movements were seen by B-mode scan. Colour Doppler was applied to visualise the tracheal fluid flow, followed by spectral Doppler to record the velocity waveforms. The records of 53 fetuses divided into five gestational age groups (20 -23, 24-27, 28-31, 32 -35 and 36 -40 weeks of gestation) containing 40 or more continuous breathing cycles (inspiration and expiration) were considered for analysis. Only regular breathing phases were examined and the volume obtained by integration of the tracheal fluid flow displaced during fetal breathing movements was calculated. Results The intra-tracheal flow volume moved during inspiration (Vi) and expiration (Ve) increased until 36 weeks of gestation after which there was a flattening until term. This suggests either a reduction of lung liquid production or a diminished lung liquid volume. The median difference between Vi and Ve was positive in the first four age groups and negative in the last one suggesting that, in mature fetuses, the effect of fetal breathing movements no longer results in an influx. Conclusions Our data demonstrate a modification in fetal behaviour that manifests itself during the last four weeks before birth and has the potential to reduce lung liquid volume.