In Vitro and In Vivo Metabolite Profiling and Identification
Assay System: 
  • In vitro: Liver microsomes, S9 fractions, or hepatocytes.
  • In vivo: Biological fluids (e.g. plasma, urine) or tissues (e.g. brain, liver) from animal studies.
  • Identification of metabolites of test compound generated in Phase I and/or Phase II metabolic reactions.
  • Determination of tentative structures of metabolites using advanced LC-MS/MS analysis.

Assay Conditions:

Test Compound Concentration           1-50 µM
Incubation Time           0.5-4 hours
pH           7.4
Temperature           37 °C
Analysis           LC-MS/MS
Number of Repeats           2

Deliverable:
 
  • Tier 1 analysis: A list of identified metabolites, including corresponding molecular masses and relative abundance of metabolites estimated from integration of peak areas in HPLC-UV/LC-MS/MS chromatograms.
  • Tier 2 analysis: Tier 1 analysis results, plus MS/MS spectra of identified metabolites, suggested metabolite structures, and possible metabolic pathways of test compound.
Test Compound Requirement: 1-2 mg dry powder or 100-200 µL of 10 mM DMSO stock solution.