Metabolite Profiling and Identification

Creative Biolabs has worked extensively on a wide variety of metabolism studies. As part of our in vitro metabolism service, we provide high-quality metabolite profiling and identification service to characterize a drug's metabolites.

The process of metabolites profiling and identification of therapeutic compounds in the early stage of drug discovery has great value in interpreting a drug's pharmacology, pharmacokinetic and toxicology characteristics and may have directive function in the following clinical tests. As we know, after a drug entering the body for a certain time, it will be eliminated either by excretion or by metabolizing to one or more metabolites. Those drugs undergoing metabolism may produce unexpected effects or potential toxicities. Therefore, it is important to identify the metabolites of a drug. Also, the knowledge of metabolic liability is useful for guiding lead optimization to overcome issues such as stability.

The FDA Guidance for Industry on Safety Testing of Drug Metabolites (2008) highlights in vitro metabolite assessment for humans and animals. Those metabolites which are unique to humans or have disproportionate levels in humans may require additional toxicological studies.

Creative Biolabs provides a variety of matrices for metabolite profiling and identification, including liver microsomal incubations, hepatocyte incubations, S9 incubations, recombinant enzymes as well as other in vivo approaches. All matrices are available from multiple species for cross-species metabolite profiling. We measure compound degradation, isolate of metabolites and elucidate the chemical structure by a variety of methods, of which the high-resolution mass spectrometry (HR-MS) is the major tool for metabolite identification both qualitatively and quantitatively. We are also able to detect radiolabeled chemicals for analysis of different types of chemistry.

Figure 1. General scheme of HR-MS-based approaches to drug metabolite detection and identification (Zhu et al. 2011).

A Typical Protocol

• A 10 µM solution of each compound is prepared in DMSO.
• Test compound is incubated with pooled liver microsomes (1 mg/mL) (or other samples, like S9, hepatocyte, recombinant enzymes) and NADPH-generating system in phosphate buffer (pH 7.4) for a period of 0 or 90 minutes at 37°C.
• Microsomes-compound matrix is quenched by addition of acetonitrile.
• After centrifugation, supernatants are transferred to new tubes, and analyzed by high-resolution mass spectrometry.
• Control samples are quenched at time zero.

The Advantage of Our High-Resolution Mass Spectrometry (HR-MS) Platform

Mass spectrometry plays a key role in drug metabolite identification and profiling. Modern high-resolution mass spectrometry (HR-MS) instruments are the development of conventional MS instruments with improved accuracy and stability, along with new data processing techniques. Creative Biolabs equipped with the most advance HR-MS equipment which has improved the quality and productivity of metabolite identification processes. The resolution is > 10,000 at full-width at half-maximum and the mass is > 5 ppm deviation.

Our advantages:

• Able to detect both primary and secondary excretory metabolites
• Various HR-MS-based analytical strategies to ensure complete metabolites identification
• Abe to study slowly metabolized drugs
• The most predictive data processing strategies

For more detailed information, please feel free to contact us or directly sent us an inquiry.

References

1. Zhu M, Zhang H and Humphreys W G (2011). “Drug metabolite profiling and identification by high-resolution mass spectrometry”. J Biol Chem 286(29):25419-25425. doi: 10.1074/jbc.R110.200055
2. Joachim D P, Kristin K I (2016). “High-resolution mass spectrometry: basic principles for using exact mass and mass defect for discovery analysis of organic molecules in blood, breath, urine and environmental media.” Journal of Breath Research 10 (1): 012001

For Research Use Only.