Evaluation of homogenization techniques for the preparation of mouse tissue samples to support drug discovery

Bioanalysis. 2011 Sep;3(17):1923-33. doi: 10.4155/bio.11.181.

Abstract

Background: In early drug-discovery research, understanding the tissue distribution of drug at the site of action can help to predict the toxicity, efficacy and exposure level of the drug. The bottleneck of tissue analysis by LC-MS/MS is the time-consuming homogenization step.

Results: Both mechanical and enzymatic techniques for mouse tissue homogenization were evaluated, which included bead beater, polytron and enzymatic digestion. Brain, bone marrow, kidney, spleen and liver tissues can be homogenized effectively using the bead beater alone. Lung and heart tissues were best treated with collagenase first and then homogenized by the bead beater.

Conclusion: Homogenization conditions for seven mouse tissues have been evaluated and optimized. These findings will expedite the preparation of tissue samples for analysis.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Chromatography, Liquid / instrumentation*
  • Chromatography, Liquid / methods*
  • Drug Discovery*
  • Female
  • Mice
  • Molecular Structure
  • Particle Size
  • Pyrazoles / chemistry
  • Pyrazoles / pharmacokinetics
  • Pyridazines / chemistry
  • Pyridazines / pharmacokinetics
  • Tandem Mass Spectrometry / instrumentation*
  • Tandem Mass Spectrometry / methods*
  • Tissue Distribution
  • Tissue Extracts / analysis*

Substances

  • N-(4-(3-(1-ethyl-3-fluoropiperidine-4-ylamino)-1H-pyrazolo(3,4-b)pyridin-4-yloxy)-3-fluorophenyl)-2-(4-fluorophenyl)-3-oxo-2,3-dihydropyridazine-4-carboxamide
  • Pyrazoles
  • Pyridazines
  • Tissue Extracts