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The following message was posted to: PharmPK
Dear Group,
We are doing tissue distribution study in rodents for our NCE. We are
homogenising the different organs in appropriate solution. Then the
homogenate obtained is centrifuged and a clear supernetant is processed
through SPE and injected in to the HPLC to know the actual levels of
NCE in
the organ.
Here I want to know how we will ensure that there is no more drug
left over
in the homogenate residue after centrifugation.
waiting for your comments on this query.
regards
Dr. Amol A. Raje
Wockhardt Research Centre
Aurangabad, India.
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The following message was posted to: PharmPK
Dear Amol,
I assume the simplest way would be to re-homogenise the residue and
process the supernatant again by SPE to inject onto the HPLC. Here I
assume that the "appropriate solution" that you have used brings out the
NCE from the tissue.
The best option to my knowledge would be LLE, however I assume that you
opted for SPE because LLE did not work, good luck, Jagannath
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Dear Amol
You need to optimize the number of homogenisations. It may be likely
that all the drug may not come out at once. So, collect the
supernatant of first homogenisation. Then, in the residue you again
add the solvent and homogenise and collect the supernatant. Combine
the supernatants. Then proceed for SPE.
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The following message was posted to: PharmPK
I have to re-extract again the pellet and check for the remaining
drug. Of course you have to be sure of your extraction method.
Dr Roberto Conti
Sigma-Tau
Dep. of Endocrinology and Metabolism
e-mail. roberto.conti.-at-.sigma-tau.it
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Dear Amol,
You will come to know this indirectly by doing some recovery studies
by spiking your compound (at concentration range expected) in a organ
homogenates of control animals and extract in the same way as that of
test samples.
Best wishes,
T. Thanga Mariappan, Ph.D.
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If LCMS you can use a labelled isotope (stable), Easiest way is to
use radiolabelled but do not chromatograph, just count for recovery.
If you cannot use either, select a compound of simliar structure and
properties, see advacned chem development
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