ABSTRACT
Many currently used thin-layer chromatographic methods for phospholipid assay rely on charring the developed plate in the presence of cupric acetate. Saturated acyl phospholipids do not react. We find that substitution of cupric sulfate results in detection of both saturated and unsaturated phospholipids. By exploiting the difference with the two reagents, one can separately estimate the amounts of saturated phospholipid. The method described here is reproducible, and we illustrate its use in determinations of (3-sn-phosphatidyl)cholines (lecithins) in amniotic fluid from problem pregnancies.
Subject(s)
Amniotic Fluid/analysis , Fetal Diseases/metabolism , Organometallic Compounds , Phosphatidylcholines/analysis , Chromatography, Thin Layer , Copper , Copper Sulfate , Female , Gestational Age , Humans , Infant, Newborn , Pregnancy , Prenatal Diagnosis , Respiratory Distress Syndrome, Newborn/diagnosis , Sphingomyelins/analysisABSTRACT
In studies of hydrolysis and synthesis of cholesterol esters by aortic enzymes, conventional methods (aside from isotope methodology) are too insensitive to measure reaction products when few aortas are used. High-performance thin-layer chromatography coupled with spectrodensitometry of plates charred after spraying with a cupric acetate-phosphoric acid reagent permitted quantitation of 10-ng amounts of cholesterol, cholesterol oleate oleate, and oleic acid. Linear calibration curves were obtained after spectrodensitometry of chromatograms containing 10-200 ng of lipid. The results were verified by radioassay.
