Glycolipid liquid chromatography

Lampio, A. and Finne, J., Sugar analysis of glycoproteins and glycolipids after methanolysis by high-performance liquid chromatography with pulsed amperometric detection, Anal. Biochem., 197, 132, 1991. 

In work on enzymic deacetylation, a series of D-glucose derivatives acetylated at specific positions was prepared, and the O-trimethylsilyl derivatives of these D-glucose acetates were separated by gas-liquid chromatography.352,353 Research on bacterial glycolipids required the characterization of 3- and 6-O-palmitoyl-D-glucose, the separation of which, as their O-trimethylsilyl and acetyl derivatives, was studied.354... 

The occurrence of 3-deoxy-D-manno-octulosonic acid in lipopoly-saccharides has prompted its synthesis,537 together with the D-gal-octo158 and the D-gluco analogs.157 Although gas-liquid chromatography was successfully used to analyze the products of these syntheses, it has been reported that methanolysis of a bacterial endotoxin lipopolysaccharide failed to yield 3-deoxy-D-manno-octulosonic acid, presumably because of the lability of the latter to acid.381 However, Kasai and Nowotny have reported four peaks for the O-trimethylsilyl derivatives of 3-deoxy-D-manno-octulosonic acid obtained by methanolysis of the glycolipid from a Salmonella minnesota mutant.538 Reduced 3-deoxy-D-manno-octulosonic acid and its methyl ester have also been analyzed successfully as their acetates.339,539... 

Liquid chromatography (l.c.) under elevated pressure has now been introduced into glycosphingolipid separation. Glycolipids are analyzed as perbenzoylated,112-116 N-acetylated O-benzoylated,"7 "8 and N-(p-nitro-benzoyl)ated O-acetylated119-122 derivatives, as well as in their native... 

Preparative and Analytical High Performance Liquid Chromatography of Glycolipids... 

Suzuki, A. Handa, S, Yamakawa, Y. Separation of molecular species of higher glycolipids by high performance liquid chromatography of their 0-acetyl-N-p-nitrobenzoyl derivatives. 

In this manuscript, we will first describe the newly developed high performance liquid chromatography of cerebroside, sulfatide, and other minor galactolipids. This method allows complete analysis of a very small amount of these glycolipids in cell or membrane preparations. This will be followed by a description of our new method of determining surface galactolipids and its application to myelin cerebrosides. 

Sialic acid was partially-purified from lipid extracts after hydrolysis in 0.05NHC1 (total sialic acid) or without hydrolysis (free sialic acid) as described in Methods. The amounts of gly-cosidically-bound sialic acid (total minus free) and free sialic acid were determined by the TBA assay. The hydrolysed samples were also analyzed using gas liquid chromatography by the method of Yu and Ledeen (15) and shown to possess essentially identical amounts of total sialic acid. However, the crude lipid extracts were too impure to make GLC the method of choice in the absence of extensive purification of the glycolipids. 

The following chapters in Advances in Lipid Methodology, Volume 3 (Cl) Chapter 3, Separation of phospholipid classes by high-performance liquid chromatography. W. W. Christie (C2) Chapter 6, Plant glycolipids Structure, isolation and analysis, E. Heinz. 

Otsuka, K. Yamakawa, T. The apphcation of droplet counter-current chromatography (DCC) forthe separation of acidic glycolipids. J. Biochem. (Tokyo) 1981,90,247-254. Kim, Y. Wang, T.C. Ma, Y.C. Liquid chromatography/ mass spectrometry of phosphohpids using electrospray ionization. Anal. Chem. 1994, 66, 3977. 

Sugawara, T and T. Miyazawa. Separation and detection of glycolipids from edible plant sources by high-performance liquid chromatography and evaporative light scattering detection. Lipids 1999,34 1231-1237. 

Snada, S., Uchida, Y., Anraku, Y, Izawa, A., Iwamori, M., and Nagai, Y., Analysis of ceramide and monohexaosyl glycolipid derivatives by high-performance liquid chromatography and its application to the determination of the molecular species in tissues, J. Chromatogr., 400, 223-231, 1987. 

Of the many analytical techniques now available to the lipid chemist, mass spectrometry (MS), is probably the one that has experienced the fastest growth in the last two decades. This is due both to the development of new techniques (gas and liquid chromatography combined with MS, soft-ionization MS, field desorption MS, atmospheric pressure MS etc.) and to the refinement of more traditional methods and their successful application to very complex problems, e.g. the elucidation of glycolipid structure, or the study of structures in lipid mixtures. Much progress has been made since the pioneering work of Ryhage and Stenhagen (1963) on fatty acid methyl esters. 

The glycosidic bond of glycolipids is readily hydrolyzed by acid, and the carbohydrates liberated can be analyzed by instrumental methods, e.g., gas chromatography (GC), liquid chromatography (LC), or colorimetric procedures can be used. In one colorimetric procedure, the total amount of hexose units in a glycolipid can be quantified by reaction with anthrone in concentrated sulfuric acid and determination by the absorbance at 635 nm. 

Analytical methods for phospholipid and glycolipid classes by normal-phase high-performance liquid chromatography (HPLC) have been incompletely reviewed. Silica columns with evaporative light scattering detection (ELSD) were mostly used. The poly(vinyl alcohol)-grafted silica column with ELSD could be used to separate them, simultaneously. 

Smith, LA. Norman, HA. Cho, S.H. Thompson, GA. Jr. Isolation and quantitative analysis of phosphatidylglycerol and glycolipid molecular species using reversed-phase high-performance liquid chromatography with flame ionization detection, J.Chromatogr., 1985,346, 291-299. 

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