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Clinical Chemistry 18: 956-960, 1972;
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Clinical Chemistry, Vol 18, 956-960, Copyright © 1972 by the American Association for Clinical Chemistry

Modified Method for Estimating the Phosphatidyl Choline:Sphingomyelin Ratio in Amniotic Fluid, and Its Use in the Assessment of Fetal Lung Maturity

Laszlo Sarkozi 1, Hanna N. Kovacs 1, Howard A. Fox 1, and Thomas Kerenyi 1

1 Departments of Chemistry, Pediatrics, and Obstetrics & Gynecology, The Mount Sinai Hospital and The Mount Sinai School of Medicine of The City University of New York, New York, N.Y. 10029.

Phospholipids of amniotic fluid were extracted, separated by thin-layer chromatography, detected with acidified ammonium sulfate, and charred; the phosphatidyl choline and sphingomyelin fractions were quantitated by transmission densitometry. The densities were recorded as integrator trace units. We used commercially available precoated plates and a solvent system to eliminate the interference of phosphatidyl serine. The procedure used for visualization eliminated the irritating and corrosive effects of the 50% sulfuric acid spray used in other methods. Omission of the acetone precipitation step decreased assay time without loss of accuracy. In 117 amniotic fluid samples obtained before delivery of infants who subsequently did not develop respiratory distress syndrome (RDS) the geometric mean density ratio of phosphatidyl choline: sphingomyelin was 3.1, range 1.8 to 8.6. In three pre-delivery amniotic fluid samples, density ratios of less than 1.5 were found. Those infants developed RDS.


Key Words: thin-layer chromatography • densitometry • respiratory distress syndrome • pulmonary surfactant

Submitted on March 24, 1972
Accepted on May 14, 1972






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Copyright © 1972 by the American Association for Clinical Chemistry.