I need to separate sugars and phenolic compounds present in a crude extract (from a fruit)
It is hard to separate phenolic compounds from simple sugars when you are working with a crude extract. I have performed a similar work with different fruits and without a previous extraction you will find lots of sugars in your chromatogram plus sugar moieties attached to Phenolic compounds. You need to choose a column that is suitable for phenolic separation and maybe you won't have co-eluent peaks.
Try ion exchange column chromatography (Strong Anion eXchange, SAX column). The sugars should elute while the phenolic compounds are captured on the solumn.
For more information, please see:
http://www.isco.com/WebProductFiles/Applications/101/Poster_and_Paper_Reprints/Purification_of_Phenolic_Flavonoids.pdf
also
http://www.isco.com/WebProductFiles/Applications/101/Application_Notes/AN87_RediSep%20Rf_SAX_Column_Applications.pdf
@PhillipMorris I would greatly appreciate if you can send me the protocol ! thanks
You can try poliamide coulmn chromatography using with water:MeOH mixture. You can start with water. It is efficent method to separate phenolics and non-phenolics
@Phillip Morris: I would appreciate if you could me your protocol as well?
A solid phase extraction column chromatography can be used for the removal of sugars from fruit extract using Bond Elut ENV cartridge (Agilent). The Bond Elut ENV cartridge is to be conditioned with 2x4ml 100% methanol, and then equilibrated with 2x4ml distilled water. Typically, 1g freeze-dried fruit extract dissolved in 4 mL MilliQ water, loaded onto cartridge, and water-soluble components eluted with 3-4 times with 4 mL of MilliQ water. The cartridge then dried out completely before elution of the non-sugar components was carried out with 3x2ml 100% methanol under slow (drop wise) flow rates.
ref. O'Kennedy N, Crosbie L, van Lieshout M, Broom JI, Webb DJ and Duttaroy AK. Effects of antiplatelet components of tomato extract on platelet function in vitro and ex vivo: a time-course cannulation study in healthy humans. Am J Clin Nutr. 2006;84(3):570-9.
Hi, I am also trying to isolate/ remove polyphenols but also would like to recover the sugars from my biomass samples! Would you be able to share the protocol for efficient separation of phenolics from sugars? Much obliged!
I also support using ion-exchange resin. Washing the sample-loaded resin with water several times must remove all the sugars while the phenolic compounds are intact, which can be further eluted with methanol.
My observation with C18 was it cleans the extract from sugars nicely. However, with sugars you might lose the water soluble phenolic acids at neutral pH. I would recommend to use acidic pH to retain the phenolic acids.
@Phillip Morris: I would appreciate if you could share me your protocol as well.
Thank you
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