PharmPK Discussion - Metabolism in human liver microsomes mediated by CYP2D6

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On 29 Apr 2002 at 10:13:00, rfernanr.at.almirallprodesfarma.com sent the message


Dear all,

I have to validate a technique to study drug metabolism in human liver
microsomes, specifically that mediated by CYP2D6. I was thinking of using
bufuralol as a probe. Could someone give me information about the assay
conditionsİsuch as  the Km value of bufuralol, microsomal protein
concentration, organic solvent and percentage used for its dissolution?

Any input will be very much appreciated

Thanks

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On 30 Apr 2002 at 13:25:33, "jignesh surendrabhai kotecha" (jskotecha1.-a-.rediffmail.com) sent the message


Hello Dear,

For in-vitro metabolism study, initially you can use drug
concentration 100µM and upto 10%DMSO in final dilution to make it
soluble.
Enzyme concentration should be around 50 pmol,and co-factor 1mg/ml
of final dilution.
Terminate the reaction using either methanol or acetonitrile ( for
100µL reaction mixture use additional 20µL of organic solvent).

warmly,
Jignesh.
jskotecha1.-at-.rediffmail.com

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