I am presently purifying some bioactive peptides from processed soybean only to realise it is contaminated with polyphenolic compounds from the bean. I will appreciate a concise answer on how these polyphenolic contaminants can be removed from the fractions.
Try extracting the polyphenols using an appropriate solvent, which you should be able to determine depending on the total polyphenol content in your peptide fraction. However, further purification using a HPLC would be better to maintain the integrity of your peptide fractions. Goodluck
You may try using hydrolysis followed by ultrafiltration and polystyrene resin separation. Attached is an article by D' Alvise et al. (2000) detailing on the procedure they used to obtain polyphenol-free alfalfa peptide.
Use reverse phase chromatography, I prefer semipreprative chromatography
You can give a try to this method:
Add 0.75 ml of buffered phenol pH 6.2 to your 1ml extract and incubate for 15 mins. Centrifuge @ 10000xg for 10 min at 4c. Collect the aqueous phase and add equal volume of ice cold acetone. Incubate at -20c for 15 min. Centrifuge for 10 min @10000xg at 4c. Collect the precipitate and wash with 80% ice cold acetone. Your peptide might be free from polyphenols.
You can give solid phase extraction (SPE) a try. It is a simple method, and there are a range of different SPE cartridges that are available such as HLB, C18 etc depending on your requirement. You can easily use these cartridges to separate the peptides from the polyphenols by a polarity-based separation. However, SPE is limited by its ability to process only small sample sizes. Depending on your sample size, you might need to use multiple cartridges. Hope this helps.
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