Dear Sir. Concerning your issue about the significance of methylation analysis for polysaccharide fractions extracted from mushroom. The polysaccharide (4 mg) was methylated according to the methods of Ciucanu and Kerek,12 and the products were isolated by partition between 5:2 CHCl3–H2O. The organic layer containing products was washed with 3 mL water three times and dried. The methylated products were then hydrolyzed with 90% HCO2H (1 mL) at 100 C for 1 h, reduced with NaBH4, acetylated with 1:1 Ac2O–pyridine and analyzed by GLC and GLC–MS using an HP-5 capillary column and the same temperature program indicated above. Quantitations were carried out from peak areas. I think the following below attached file may help you in your analysis:

Thanks

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Dr Sadia

Follow the following links

http://www.nature.com/nprot/journal/v7/n9/fig_tab/nprot.2012.081_F3.html

https://ast.uga.edu/glycosyl-linkage/

http://www.stenutz.eu/yellow/applic.html

Regards......

@ Isam Eldin Hussein Elgailani : Dear sir thanks for sharing this protocol . but my question is why methylation analysis is carried out for polysaccharide fractions ? what is the rationale behind doing this assay for these bioactive compounds.

Thanks