In Vitro MetID (Metabolite Profiling and Identification)

In vitro MetID utilizes ultra-high performance liquid chromatography (UPLC) coupled with a photodiode array detector (PDA) and high-resolution mass spectrometry (HRMS) for data acquisition, and combined with data processing software (Xcalibur, Compound Discoverer, Masslynx, Mass Metasite, etc.) and multiple data processing techniques (mass deficit filtering, background deduction, characteristic ion extraction, and isotope filtering, etc.) to search for and to identify metabolites.

When conducting in vitro MetID study, the incubation time and incubation matrixes will be adjusted based on the purpose of the experiment and the in vitro stability of the test compound in the incubation medium. For example, when performing a metabolic soft spot analysis, long incubation time is not recommended, generally 1-2 half-life are suitable. When conducting in vitro MetID study of slow metabolic compounds in hepatocytes, adherent cells or relay method would be suitable for incubation.

Metabolites which are only found in human or have significantly higher proportion in human than other species or have potential toxicity (e.g. glucuronidation metabolites) require extra attention.