Permethylation of the Sample

Methylation of a carbohydrate is most practically achieved by using the Hakomori procedure,9 which has largely replaced the Purdie, Haworth, and Kuhn methylation methods. 

Completeness of the methylation reaction is a prerequisite for successful methylation analysis. It can be directly detected from the disappearance of the infrared absorption of the hydroxyl groups,9 but this is often not possible because of the small amount of sample available. 

Use of Potassium tert-Butoxide to Generate the Methylation Reagent 

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As early as the mid-1980s, the permethylation of oligosaccharides was applied to for their analysis when employing secondary ion mass spectrometry (SIMS) and fast atom bombardment (FAB). Indeed, with the advent of ESI and MALDI a few years later, this method of sample modification was foimd to be perfectly suitable for these techniques, given the enhancement in ionization efficiency that resulted from permethylation of the sample [63]. 

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