Back to the Top
Hi all,
Can someone suggest the sample preparation strategies for developing
and validating a LC-MS method for an IND in C57 BL/6 mouse urine? I
need to perform a standard curve, and quality control type (QC)
experiments for my drug in C57 BL/6 mouse urine. We use a protein
precipitation approach for plasma using MeCN/MeOH. I want to know if
a similar approach can be used by just substituting urine for plasma
or any modifications are necessary while working with urine.
Many Thanks,
Pradeep B.Lukka
Auckland Cancer Society Research Centre
The University of Auckland
Auckland
New Zealand
Tel - 64 (0) 9 373 7599 x89845
Back to the Top
The following message was posted to: PharmPK
Dear Lukka, Hi to you and all our memberes in the grop.
Yes, you can start by try using the same method as for plasma and if
did not work look or develope new method keeping in mind doing all
the steps requirement to fillfulment the validations as its stated by
FDA or other.
Best regard and good luck.
Professor Khalil I M Al-Khamis, Ph.D
Back to the Top
The following message was posted to: PharmPK
This is the trap of the partial validation. The range, interferences
and metabolites, and stability will be different in urine compared to
plasma. The most conservative approach would be to start with your
chromatography/fragmentography method then modify to fit urine. This
may mean a complete (full) validation. You may need a
hydrolysis/glucuronidase step then an extraction step to clean up the
sample, which will need to be validated as well. The urine
concentrationwill be much higher. Unless you are controlling the
collection of urine (bladder cannula) the time course concentrations in
urine will be somewhat dissociated from the plasma profile.
Ed O'Connor, PhD
Technical Director, Immunoanalytical
Tandem Laboratories
115 Silvia Street
West Trenton, New Jersey
609-228-0243
Back to the Top
The following message was posted to: PharmPK
Dear Professor Khalil I M Al-Khamis,
why do you suggest the use of organic solvents for urine sample
preparation?
Proteins should not be present in urine, so I was wondering if there
is any advantage deriving from this approach.
Best regards,
Marco Lai
Mass Spectrometry Lab
R&D Dompe pha.r.ma S.p.A.
Via Campo di Pile, 67100 L'Aquila - Italy
Phone: +39 0862 338332
Fax: +39 0862 338219
e-mail: lai.marco.at.dompe.it
Back to the Top
The following message was posted to: PharmPK
Dear Marco Lai; HI
If I remember I did suggest replacing the plasma by the
urine .......... so on. If its different I apologize to that and what
you said absolutely right.Thanks and best regard.
Professor Khalil I M Al-Khamis, Ph.D
Back to the Top
The following message was posted to: PharmPK
There are always proteins in urine, much less than in plasma but they
are there. Higher levels indicate renal/liver/immuno issues but they
are always there.
Ed O'Connor, PhD
Technical Director, Immunoanalytical
Tandem Laboratories
115 Silvia Street
West Trenton, New Jersey
609-228-0243
Want to post a follow-up message on this topic?
If this link does not work with your browser send a follow-up message to PharmPK@boomer.org with "Method development validation in mouse urine" as the subject | Support PharmPK by using the |
Copyright 1995-2011 David W. A. Bourne (david@boomer.org)