I wanted to evaluate the exact molecular weight of chitosan (from shrimp shell; degree of deacetylation NLT 75.00% and molecular weight range 3800-20,000 Daltons) using gel permeation chromatography with dextran as a standard. So I am getting the peaks of the standard properly, but I am not getting any peaks for chitosan. Previously I tried evaluating the same in the case of kappa carrageenan and was able to get the peaks properly. But in the case of chitosan, it's not happening. What might be the reason behind it?
The column is fine, as the same column was used for carrageenan.
Standard (Dextran) is also good, as the peaks are coming.
I have attached the specification of chitosan.
Did you solubilize chitosan in acidic solution of acetic acid. it may be the reason. I suggest to measure at pah 4 or 3.5 at which it has the largest hydrodynamic radius
Dmitriy Berillo, I solubilized it in 1% glacial acetic acid. In the mobile phase, I am using H₂O, 0.1 M sodium chloride, and 0.3 vol% trifluoroacetic acid.
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