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Hello,
I am working on the compound Thioacetamide and its metabolites Thioacetamide
suof oxide, and acetamide. I am quantitaing these from the blood and liver
tissues by HPLC. These compounds are highly polar. I am employing
liquid liquid extraction using ethylacetate as a solvent and found that I am
not getting sufficient recovery. also looks like the procedure is not clean
and reproducible. I am also presently trying reverse phase solid phase
extractions. THese compounds have high solubility in ethanol followed by
water. Thioacetamide was earlier used as an antifungal agent and is no more
used pharmacologically.
I would want some input with regard to extractions from these biological
fluids for quantitation.
If you need any more information on these, i can provide for you,
looking for a reply,
shashi
961kumar.at.alpha.nlu.edu
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Shashi,
If your compounds are not highly bound to plasma proteins and if
concentrations are in ug/mL range you could try a simple
deproteination by adding 30 % of ice-could ethanol followed by
centrifugation. You could inject the supernatant directly onto the
column. There should be a pre-column in your HPLC system!
Hope this help
Vladimir
--------
Vladimir Piotrovsky, Ph.D. Fax: +32-14-605834
Janssen Research Foundation Email: vpiotrov.-a-.janbe.jnj.com
Clinical Pharmacokinetics piotrovskijv.at.jrf.be
B-2340 Beerse
Belgium
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Extracting polar compounds with ethyl acetate is not very good strategy.
Since you mentioned you metabolites are polar. I recommend using
protein precipitation by acetonitrile (1:2), MeOH (1:2), Perchloric acid
(1:0.3) after this precipitation, the chromatography of the aliquot is not
clean (some times it may workout) Subject the supernatant to a solid
phase extraction or a liquid - liquid with Methylene chloride-Amyl alcohol
(98:2). Addition of amyl alcohol for some miraculous reason improves the
extractability of very polar compounds from biological matrices.
In a lighter sense when this compound is no longer used
pharmacologically why you are wasting your time working on this
compound (Just take it easy).
Prasad Tata, Ph.D.
Otsuka America Pharmaceutical, Inc.
2440 Research Blvd.
Rockville MD 20850
>>> SHASHI KUMAR VET TOXICOLOGY by way of David_Bourne When
Hello,
I am working on the compound Thioacetamide and its metabolites
Thioacetamide suof oxide, and acetamide. I am quantitaing these from
the blood and liver tissues by HPLC. These compounds are highly polar.
I am employing liquid liquid extraction using ethylacetate as a solvent and
found that I am not getting sufficient recovery. also looks like the
procedure is not clean and reproducible. I am also presently trying
reverse phase solid phase extractions. THese compounds have high
solubility in ethanol followed by water. Thioacetamide was earlier used
as an antifungal agent and is no more used pharmacologically.
I would want some input with regard to extractions from these biological
fluids for quantitation.
If you need any more information on these, I can provide for you,
looking for a reply,
shashi
961kumar.-a-.alpha.nlu.edu
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Dear Dr. Prasad Tata,
I am writing this to inform you that although now thioacetamide is not used
pharmacologically, we are working on a concept called tissue regeneration,
in liver. I am trying to quantitate this compound and its metabolites
to prove a different point altogether, NOT NECESSARILY to be useful clinically.
I take your comments definitely in a lighter sense, and appreciate your
interest.
this is for your clarification,
Shashi Kumar Ramaiah, DVM., MS
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