Do anyone know a method for sugar extraction from cyanobacteria for HPLC analysis?
Sugar can be extracted by Methanol or Acetonitrile with water by sonication for 2-5 minutes. i am saying this as a analytical chemist. i am not aware of biological part but sugar can be extracted easily by many of the solvent polar solvent.
Please specify what sugar you want to extract sucrose, fructose, glucose, maltose or else?
Regards
Selvan
Hi Selvan,
I would like to extract the total sugars from the organism......
you can extract with water and ethanol or methanol or acetonitile, but i do not know what is organism; by sonication of your sample for 10-15 min with the above solvents may bring out your sugar as it is polar nature and the below link may help you
www.chem.agilent.com/Library/primers/Public/59883294.pdf
Regards
Selvan
Thank u selvan.....I will try sonication of my sample with acetonitrile....and thanks for the link too....
gud nite...
Dinesh
you can see this article: Rigane et al. (2011) Journal of food science. 965-973.
Hi
The suggestion of Selvan are good.
Moreover, under this link you can find a general protocol for extraction of sugar from Cyanobacteria:
http://www.springerlink.com/content/r228516l52183436/
Do not forget the composition of cell wall in cyanobacteria which will contain consistent amounts of sugars. The lipopolysaccharide mostly consist of consisting of fucose, mannose, galactose and glucose. But also other sugars such as glucose rhamnose, mannose, arabinose depending on the strain (see: http://jb.asm.org/content/182/5/1191.full). By the extraction with Ethanol the lipopolysaccharide will precipitate with the debris together the proteins and lipids.threfore, consider an additional strategy, enzymatic, for this fraction, If you are also interested of these sugars.
For a subsequent HPLC analysis you can use ion exchange columns such as Hi-Plex (Hi-Plex H: Strong cation-exchange resin consisting of sulfonated, crosslinked styrene-divinylbenzene copolymer in hydrogen form,) from Agilent (see: http://www.chem.agilent.com/Library/applications/5990-8801EN%20Hi-Plex%20Compendium.pdf) or HC-75 (Hydrogen form, Gel-type cation exchange columns) from Hamilton (http://www.hamiltoncomp.com/HPLC/applicationIndex.php).Under both links you will find also HPLC condition.
Saluti
Ignazio
@ Rigane...
Hi, can you please send a copy of the article. I couldn't find it online.
Thank u....
@ Ignazio
Hi....thank you very much....we used to extract the exopolysaccharides and analyze it... i would like to have the total sugars of the organism.....anyway thank you very much.........
Hi
you do not need the article, since in the first page the extraction pocedure is described (at the end).
Ignazio
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