Hi all,

I am trying to access the shift in molecular weight of a sulfated polysaccharide before and after hydrolysis. Nothing has worked well so far with my 2 methods:

1. Loading the products on a gel and staining with toluidine blue. This was unsuccessful as the native polysaccharide showed a much smaller MW than what it should be - perhaps because it is more negatively charged than the sulfated dextran standards.

2. HPLC - unfortunately, although I see a curve for the native product, I cannot see the curve of the hydrolysized (low molecular weight) compound. Pre-labelling my polysaccharide with toluidine blue and tracing its absorbance also didnt seem to work.

For hydrolysis, I treat with 0.1N HCl for 10m at 90˚C. Is it possible that this converts a 200-500kDa polysaccharide into a mono/ogliosaccharide, making it harder to detect??

Any insights would be highly appreciated - this is driving me nuts!

Thank you.