Metabolite Profiling and Identification in Mammalian Tissues or Fluids from In Vivo Studies
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.