Influence of phospholipid saturation on classical thin-layer chromatographic detection methods and its effect on amniotic fluid lecithin/sphingomyelin ratio determinations.

We compared seven techniques, commonly used for detection of amniotic fluid phospholipids in thin-layer chromatography, with respect to their sensitivity to saturation of the fatty acid carbon-chain of lecithin. The techniques fell into two classes: sensitive and insensitive; those classed as saturation sensitive were less than or equal to 10% as sensitive to fully saturated lecithin as to lecithin with singly unsaturated acid moieties. Color development increased with the number of carbon-carbon double bonds per molecule but required only a single unsaturated acid ester in either the alpha or the beta position. Mixtures of lecithins with defined saturation, when detected by saturation-sensitive methods, mimicked the uneven coloration of amniotic fluid lecithin bands, supporting the hypothesis that this uneven coloration results from natural saturation heterogeneity. Detection techniques representative of these classes are shown to give widely differing values for the lecithin/sphingomyelin ratio for actual specimens of amniotic fluid.