In the present study, we sought to assess the clinical efficacy of the optical density of amniotic fluid at 650 nm (A650) to predict lung maturity in the human fetus. The A650 of 113 samples of amniotic fluid obtained from 16 to 45 weeks' gestation was determined. Mature values (A650 greater than or equal to 0.11) were not observed until 35 weeks' gestation but were always present after 39 weeks' gestation. In those infants delivered within 48 hours of amniocentesis, the absence rates of respiratory distress syndrome were the same with a mature lecithin/sphingomyelin (L/S) ratio (97.9%) and a mature A650 (97.3%). However, immature values in both tests were poor prognosticators of respiratory distress syndrome, with a rate of 37.5% with the immature L/S ratio and 15.8% with the immature A650. A mature A650 may be substituted for the L/S ratio, but an immature A650 is less reliable. In addition, we found that differences in centrifugation altered the A650 value, whereas exposure to light and cold storage did not.