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Link to original content: https://pubmed.ncbi.nlm.nih.gov/1524204/
A comprehensive procedure for preparation of partially methylated alditol acetates from glycoprotein carbohydrates - PubMed Skip to main page content
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. 1992 May 15;203(1):101-8.
doi: 10.1016/0003-2697(92)90048-c.

A comprehensive procedure for preparation of partially methylated alditol acetates from glycoprotein carbohydrates

K R Anumula  1 , P B Taylor
Affiliations

A comprehensive procedure for preparation of partially methylated alditol acetates from glycoprotein carbohydrates

K R Anumula et al. Anal Biochem. .

Abstract

Various steps involved in the preparation of partially methylated alditol acetates (PMAAs) from glycoprotein-derived carbohydrates were improved to obtain the derivatives in a rapid manner with excellent yields. Carbohydrates were permethylated in dimethyl sulfoxide (DMSO), using a fine suspension of sodium hydroxide and methyl iodide (CH3I). The fine suspension of NaOH was prepared conveniently from commercially available 50% aqueous NaOH in DMSO by sonication and washing the precipitate with DMSO. Methylation of ovalbumin and fetuin glycopeptides using the fine suspension of NaOH and CH3I was complete within 5 min, and the methylation reaction did not generate any nonsugar artifacts. Methylated carbohydrates without any purification were hydrolyzed in a mixture of volatile organic acids, which permitted rapid removal of the acids from samples by evaporation. Acetylation of partially methylated alditols with acetic anhydride for 2-4 h at ambient temperature using 4-N,N'-dimethylaminopyridine as a catalyst and the reaction was free from generating nonsugar reaction artifacts. The reaction time course for methylation, hydrolysis, and acetylation was determined to obtain optimum reaction conditions for preparation of the PMAAs. The procedure facilitated rapid identification and quantitation of PMAAs due to diminished reaction artifacts and the quality of the chromatogram depended only on the purity of starting material and the reagents used for the methylation analysis. Utility of these simple methods for rapid methylation analysis was demonstrated in the characterization of oligosaccharides isolated in small amounts using a carbohydrate analyzer.

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