A new procedure for the determination of lecithin and sphingomyelin in amniotic fluid is described. rapid column chromatography in the sample preparation steps with computer-assisted infrared spectroscopy in the final determination. Lipids are extracted from amniotic fluid with a chloroform/methanol solvent. The organic layer is separated and evaporated to dryness. The residue is The new method couples the use of simple, redissolved i n chloroforn and Fassed through a hydroxyapatite column to remove interfering constituents. The complete isolation cflecithin and sphingomyelin fror other li Fidsis confimec by both infrared spectroscopy and thin layer chronatcgrayhy.