Separation of adreno-ovarian steroids by thin-layer chromatography

The major adreno-ovarian steroid hormones (progesterone, estrone, 17α-estradiol, 17β-estradiol, estriol, corticosterone, cortisone, and cortisol) have been separated simultaneously on a single TLC plate without recourse to transfer chromatography. The plate was developed successively twice in benzene/ethanol (95:5, v/v) solvent system. It was then sprayed with rhodamine 6G and a line was drawn isolating the already separated least polar and medium-polar steroids (progesterone, estrone, 17α-estradiol, and 17β-estradiol) with the help of ultraviolet light. Then 5 ml methanol per 100 ml solvent in the tank was added and the plate again developed 2–3 times up to the line drawn, when polar steroids (corticosterone, cortisone, cortisol, and estriol) separated out.     

Separation of sterols by vapor-programmed thin-layer chromatography

By vapor-programmed thin-layer chromatography on silica gel, it was possible to separate cholestanol from cholesterol and stigmastanol from beta-sitosterol. The method was applied for the analysis of beta-sitosterol-3-(14)C.     

Separation of glycoprotein-derived oligosaccharides by thin-layer chromatography.

A thin-layer chromatography (tlc) system has been developed for the separation of glycoprotein-derivedoligosaccharides. The method involves chromatography on silica gel using n-propanol/acetic acid/water (3:3:2 v/v) as the solvent. This tlc method was used to separate pathological oligosaccharides isolated from individuals with G_M1 gangliosidosis and with neuraminidase deficiency. The results indicate the potential usefulness of the system in the analysis of complex carbohydrates.     

Separation of neutral glycosphingolipids and sulfatides by thin-layer chromatography

Two one-dimensional systems for separation of glycolipids from total lipid extracts of tissues by thin-layer chromatography are described. System I used, as adsorbent, an alkaline mixture of silica gel without CaSO(4) binder (75%) and magnesium silicate (25%), and the lipids were "developed" with three successive solvent mixtures. The separated compounds (from the fastest to the slowest moving) were: ceramide, ceramide monohexosides, sulfatides, ceramide dihexosides, psychosine, ceramide trihexosides, and ceramide N-acetylhexosamine trihexosides. In system II a two-step development was used on an adsorbent consisting of silica gel without CaSO(4) binder (80%) and magnesium silicate (20%). The separated compounds were: ceramides, ceramide monohexosides, and ceramide dihexosides. Psychosine and sulfatides as well as ceramide trihexosides and ceramide N-acetylhexosamine trihexosides were not separated. In both systems all neutral lipids moved to the very top of the chromatogram and phospholipids stayed at the origin. Application of systems I and II for separation of glycolipids was demonstrated on total lipid extracts from animal tissues.     

Separation of bile acids of rat bile by thin-layer chromatography

The common bile acids of rat bile (chenodeoxycholic, hyodeoxycholic, cholic, alpha-muricholic, and beta-muricholic acids) are completely separated by a new thin-layer chromatographic system.     

Separation of sterol acetates by column and thin-layer argentation chromatography

Column and thin-layer chromatographic systems employing silver nitrate-impregnated adsorbents are described for the separation of sterol acetates which differ in the number of double bonds in the steroid nucleus or side chain.     

Separation of phosphoinositides and other phospholipids by two-dimensional thin-layer chromatography

A simple, rapid, two-dimensional TLC system is presented which resolves the four phosphoinositide cycle phospholipids as well as all commonly encountered major and minor phospholipids. Ca2+-free lipid samples are loaded onto silica gel HL plates and developed first in 48:40:7:5 chloroform:methanol:water:concentrated ammonia, and then in 55:25:5 chloroform:methanol:formic acid. The method was applied successfully to human erythrocytes, human platelets, and BL/VL3 murine lymphoma cells.     

Separation of gluco- and galactocerebrosides by means of borate thin-layer chromatography

Gluco- and galactocerebrosides can be separated by thin-layer chromatography on Silica Gel G prepared with sodium borate solution instead of water. The most successful developing system was chloroform-methanol-water-15 M NH(4)OH 280:70:6:1.     

Separation of phosphatidylinositols and other phospholipids by two-step one-dimensional thin-layer chromatography

A quick and efficient thin-layer chromatographic procedure is described for the separation of phosphatidylinositol-4,5-bisphosphate, phosphatidylinositol-4-monophosphate, phosphatidylinositol, phosphatidylcholine, phosphatidic acid, and phosphatidylethanolamine. The method involves development of the phospholipids successively in two different solvent systems but in the same direction. The method is simple, reproducible, and gives good resolution of the six different phospholipids tested. About 8-10 32P-labeled phospholipids isolated from rat hepatocytes were separated by this method; the six mentioned above were identified. Thus, the technique has potential application for both qualitative and quantitative analysis of phospholipid mixtures, such as the phosphoinositides, in cell or tissue extracts.