Thin-layer chromatography glycolipid

It has been demonstrated that ILMs are suitable for qualitative and quantitative analyses of low-molecular weight compounds of biological interest, for example, carbohydrates, vitamins and amino acids [38], and glycolipids [40]. ILMs were further used for fhe direct analysis of alkaloids, anesthetics and antibiotics, separated by thin-layer chromatography (TLC) [46]. For this purpose, the ILM was spotted onto the fractions on the TLC-plates and the complete plate was measured in MALDI MS without the need for additional pretreatment of the TLC-samples. The mass deviation inherently caused by the inhomogeneous surface of fhe TLC-plafe was balanced by using the... 

Svennerholm, E. Svennerholm, L. The separation of neutral blood-serum glycolipids by thin-layer chromatography. 

The neutral lipid fraction from the DEAE-Sephadex A-50 column was combined with the lower phase obtained after Folch partition of the total lipid extract and the combined lipids dried. To the same flask, 10Q ml of 0.6 M NaOH in methanol was added. The mixture was incubated at 37°C for 5 hours. Five volumes of acetone were then added and stored overnight at 4°C. The precipitate was collected by centrifugation at 4°C and dissolved in C M (4 1, v/v). After application to the column (2.0 x 25 cm), the column was washed with chloroform. Neutral glycolipids were then eluted with tetrahydrofuran H2O (10 1). Fractions containing neutral glycosphingolipids were pooled and their glycolipid content examined by thin-layer chromatography. 

Figure 1. Thin-layer chromatography of the glycolipids with Forssman activity...

Glycolipid analysis. Gangliosides were extracted with chloroform-methanol (2 1), purified on Sephadex G-25 columns and separated into individual species by thin-layer chromatography as described previously (12). 

Thin Layer Chromatography. Two dimensional preparative TI was used to purify Thy-1 glycolipid. All experiments were performed using Silica Gel 60 TLC plates (E. Merck, Darmstadt, West... 

This problem was approached by incorporating radioactive percur-sors into the glycolipids of both brain and lymphoma cells of the Thy-1.2 and 1.1 types. We have used C-palmitate as a per-cursor of ceramide, and - C-N-acetylmannosamine as a percursor of sialic acid (7). Glycolipids were isolated and the radioactive gangliosides were resolved by two-dimensional thin layer chromatography in two different solvent systems followed by autoradiography. 

PGL-I, homogeneous by thin-layer chromatography, through reverse-phase HPLC showed heterogeneity based on fatty acid content, and PD-MS analysis demonstrated the existence of the C3o,C3o mycocerosate glycolipid, the C32,C32 species, and the species.27 Two other minor phenolic glycolipids isolated from M. 

The homogeneity of glycolipids separated by the various procedures already described should be determined by thin-layer chromatography or HPTLC in several solvent systems. Mixtures of chloroform/methanol/water in different ratios, such as 60 35 8 v/v or 55 45 10 v/v as well as those with added calcium (chloroform/methanol/0.02% CaCl2.2H20, 60 40 9 v/v) or ammonia (chloroform/methanol/15M ammonia/water 60 35 1 7 v/v ) are particularly useful for the separation of gangliosides.Neutral and acetylated glycolipids are separated by chloroform/methanol/water (60 25 4 v/v) and 1,2-dichloromethane (DCE)/methanol/water (88 12 0.1 v/v) respectively. 

The lipids were separated on a high-performance thin-layer chromatography (HPTLC) plate. The developing solvent was a mixture of chloroform, methanol, and 0.2% aqueous CaCl2. Orcinol reagent and Dittmer s reagent were used to detect glycolipids and phospholipids. 

Phospholipids and glycolipids have been usually fractionated from total lipids by thin-layer chromatography (TLC) or column chromatography. Then, they were later analyzed by HPLC separately. 

Fuchs, B. 2012. Analysis of phospoUpids and glycolipids by thin-layer chromatography-matrix-assisted laser desorption and ionization mass spectrometry. J. Chromatogr. A, 1259 62-73. 

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