Hi there!
I've recently synthesised a 34-amino acid polypetide using Fmoc chemistry and I am having problem purifying it. My HPLC chromatogram contains many peaks (mostly byproducts) and as I have identified the peak purifying it out is proving to be very difficult. The HPLC I use have as fractionator but I have to now collect it manually because the peaks are so close to each other. Eventhough I have acquired the proper peak, the MALDI mass spectrum analysis still indicate a few peaks present and it is not 99% pure. Do you have any suggestion as to how I can improve my purifcation method?
Parameters:
C18-column
Buffer A: 100% MQ water, 0.1% TFA
Buffer B: 80% MeCN, 20% MQ water, 0.08% water
Peptide elutes at 47% B.
Chromatogram is shown below.
Hoping to hear from you
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