We are working on dextran quantification in urine samples using HPLC-MS. Our first step is dextran hydrolysis using a dextranase (EC 3.2.1.11) from Penicillium sp.
We need a method to control whether the hydrolysis step took place correctly, so we thought about adding some dextran derivates and simultaneously detecting their fragment ions. I was unable to find any data about the effectiveness of our dextranase regarding DEAE-dextran or dextran sulfate which, after hydrolysis, should yield compounds easy to detect and distinguish from products of dextran hydrolysis.
Have you ever had any experience with those reactions?
Thank you for your answers!
Sorry we cannot help you, Dextran and compounds are not within our list of products.
Best regards
Jens Kraeber
Provided that the degree of substitution is not too great it should reduce the molecular weight of your dextrans. However, when a branch point or substituent is close by it will give an oligosaccharide - the rest will be glucose.
Urine will normally contain some glucose, so its presence is to be expected.
I would not expect any dextran to pass the kidneys because of the size.
Dear prof. Stenutz,
thank you for your response. I also think that when the degree of branching (or substitution) is not very high it should not be a problem, but dextran sulphate is rather highly substituted and that is why I ask for any personal experience.
As far as i know dextran with molecular weight up to 70 kDa is quite rapidly excreted from the body with urine (Clinical Pharmacokinetics 12: 123-135 (1987))
You might be better off using a chemical degradation, e.g. acid hydrolysis, since this would give you a more predictable result. If you get oligosaccharides from enzymatic hydrolysis they may differ in subtle way and give separate peaks in HPLC.
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