PharmPK Discussion - Red blood cell partition - technical advice

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On 20 Dec 2006 at 18:55:22, "Tarra Fuchs" (tt1_chemistry.at.hotmail.com) sent the message


The following message was posted to: PharmPK

I recently tried following a protocol to determine the RBC partition
in vitro.  This procedure included obtaining fresh whole blood
(collected with anti-coagulant), dosing the blood and incubating for
1 hour. After the incubation, a portion is kept aside (and diluted)
to represent the whole blood component.  The rest of the whole blood
is centrifuged. The plasma component is collected/diluted as is the
RBC component.

I had a very difficult time pipeting the RBC component. It seems to
have coagulated despite the anti-coagulant.  I am not sure if the
problem is that there was not enough anti-coagulant, the sample needs
to be warmed back up to 37C or if I should try using a different kind
of pipet tip ( I am working with rodent plasma at the moment, so my
volumes are < 1mL)

The procedure I have calls for the use of tritiated water, to account
for volume loss due to pipeting of the blood, but the pipeting error
seems so large right now, that I am uncomfortable with the experiment.

Any suggestions would be appreciated.

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