One hundred forty-one amniotic fluid samples were analyzed for disaturated phosphatidylcholine (DSPC), phosphatidylglycerol (PG) and the lecithin/sphingomyelin (L/S) ratio. The L/S ratio was measured by two-dimensional thin layer chromatography. Mature levels (positive) were defined as an L/S ratio of 2:1 or more, a DSPC level of 100 µg/ml or more and a detectable level of PG. The DSPC value agreed with the L/S ratio in 114 samples (80.9%). Fourteen of 17 infants with respiratory distress syndrome (RDS) (82.4%) had immature L/S ratios and immature DSPC levels. RDS developed in 16 of 40 (40%) children with immature L/S ratios, and in 15 of 33 (45.5%) children with immature DSPC levels. The true-positive rates of the L/S ratio and DSPC levels were 99.0 and 98.1 %, respectively. PG had a low true-negative rate, as only 17 of 67 (25.4%) samples without detectable levels of PG were associated with RDS. However, when PG was present, it had a 100% predictive value of no-RDS. In conclusion, DSPC is nearly equal to the L/S ratio measured by two-dimensional as concerns diagnostic accuracy. PG is useful as an additional index for predicting lung maturation.