Quantitative Estimation of Some Phosphatides and Their Hydrolysis Products by Thin Layer Chromatography.

Abstract

Many improvements and simplifications have been made in the analysis of phosphatides and water soluble phosphate esters of glycerol. Silicic acid impregnated paper techniques have greatly speeded up the process of identifying and quantitating phosphatides ( 1,2,3). Dawson and coworkers (4,s) have carried out extensive studies on paper of the hydrolysis products of phosphatides. With the introduction of TLC,* qualitative separation of lipids and phospholipids was speeded up remarkably. The precision of the separations offered the possibility of quantitating the compounds so separated on thin layer plates. Two groups have analyzed some phosphatides quantitatively by TLC (6,7). The separation of the hydrolysis products of phosphatides on TLC has also been reported recently (8,9) ; however the separations achieved were not suitable for quantitation.In this report we describe a procedure for quantitative analysis of phosphatides and of the water soluble choline and ethanolamine * The following abbreviations are used: TLC, thin layer chromatogram or

Lipids of Human Testes Removed at Orchidectomy

“…Aliquots of total lipids obtained by extraction with Folch reagent were used for determinations of phospholipid phosphorus (Doizaki & Zieve, 1963), triglycérides (Sardesai & Manning, 1968), and free and esterified cholesterol (Sperry & Webb, 1950), after separation by thin-layer chromatography on Adsorbosil 2 (Applied Science Laboratories) using the solvent system petroleum ether (80 vols), diethyl ether (20 vols) and glacial acetic acid (1 vol.). The factor 25 was used to convert mg to mg phosphatides.…”

“…or on glass plates coated with Silica Gel HR (product of Merck and Co., Darmstadt, Germany, supplied by Applied Science Laboratories, State College, Pa.), using the one-69 dimensional system: chloroform/methanol/28% ammonia (65/25/4 by vol.). After developing and charring lightly with sulphuric acid, the identified spots were scraped quantitatively into test-tubes and phosphorus was quantified by the method of Doizaki & Zieve (1963). All analyses of lipid fractions were carried out at least in duplicate.…”

Phosphoglycerides and Phospholipases

“…Thin-layer ihromato~;raphy.--Since the review by Mangold (28) on the separation of phospholipids by thin-layer chromatography, this technique has been the subject of numerous papers (29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40). Excellent results have been obtained, particularly by two-dimensional techniques.…”

Alterations in pulmonary surface active lipids during exposure to increased oxygen tension.

“…(14). Since fraction 4 was found to be more than 90%o lecithin, samples of this fraction were deacylated by alkaline methanolysis (15) and fatty acid methyl esters determined by gas-liquid chromatography.…”