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I'm using cryopreserved hepatocytes for in vitro measurement of drug
intrinsic clearances with substrate depletion method. Samples are to
be analyzed with LCMS. Can someone kindly share with me the matrix
where the standard curve samples are spiked in? Idealy, it seems it
should be the same hepatocytes, but it seems costly to use
cryopreserved hepatocyte just to make standard curve samples and it
seems some kind of inactivation thing need to be done first to
inactivate the enzymes. So is there any alternative, e.g. culture
medium or liver tissue homogenate? Thanks!
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The following message was posted to: PharmPK
You can try rat hepatocytes just by producing one in the lab and
inactivate them (you can
get lot of cells with just one rat)
Hope this will be an alternate for human hepatocytes.
thanks
sripal
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The following message was posted to: PharmPK
Changing of the matrix for standard curve is usually
not suggested..Because, once you change different
matrix for your standard curve preparation, protein
binding and hence extraction recovery of drug gets
affected depending on the case..Usually you can add
hepatocytes to a sample already having terminating
solvent so as to inactivate it..and then you can
proceed with your standard curve preparation..
Regards
SK. BASHA
SK. Jafar Sadik Basha
Junior Scientist
Drug Metabolism and Pharmacokinetics,
Discovery Research,
Dr.Reddy's Research Foundation,
Bollaram Road, Miyapur, Hyderabad-49
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