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Alternatives to Laboratory Animals - ATLA

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Reduction of animal usage by serial bleeding of mice for pharmacokinetic studies: application of robotic sample preparation and fast liquid chromatography-mass spectrometry.

Bateman, K.P., Castonguay, G., Xu, L.J., Rowland, S., Nicoll-Griffith, D.A., Kelly, N. and Chan, C.C.

Journal of Chromatography B, 754(1), 245-251 (2001).

Typically. pharmacokinetic studies in mice require one animal per time point, thus resulting in differences due to dosing error. animal to animal variation and more importantly the euthanasia of a large number of animals. A method for the determination of pharmacokinetic data from serially bled mice to support early drug discovery is described. Sample analysis relies on liquid chromatography coupled with tandem mass spectrometry permitting robust and reproducible analysis requiring approximately 3 min per sample. Several parameters are discussed including the method of sample collection, preparation and analysis. The use of serially bled mice has lead to a remarkable reduction in animal usage and a corresponding reduction in compound required for such experiments. Using conventional methodology. a nine-point pharmacokinetic curve with four animals per time point would require 36 mice. With the method described below. only four mice in total are used and euthanasia is not required, permitting reuse after several weeks recovery and washout. Also. pharmacodynamic-pharmacokinetic correlation is possible and is demonstrated using a mouse model of diabetes.