cleavage of glucuronide from epicatechins

Waz wazungy at
Tue Apr 15 22:12:23 EST 2003

Currently I am trying to cleave the matabolites of several different
epicatechins to get the free compound.
I have used crude helix pomatia to effect this cleavage, but the reaction
seems to also destroy an ester bond in the target molecules as I get a rise
and fall in the target analytes when icnubating over a period of time (Plot
incubation time vs analyte signal).
I suspect that the crude glucuronidase (which is known to have sulfatase
activity) may also have esterase properties. I have found that purified
glucuronidase from this species does not perform as well as the crude. If a
co-factor is missingm what would it be??

I have tried several acetylcholine and choline-esterase inhibitors at
various strengths, but to no effect. I feel these are too specific to their
substrates. The best I can do is slow the rate of analyte degradation.
Control samples without the glucuronidase are far much more stable.

What is it in this crude glucuronidase (sigma helix pomatia) that cleaves
ester groups (our suspicion), and what can I use to block this action?

Any thoughts our guidance to proper sights would be well appreciated.

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