Availability of ADME data as early as possible during drug development is of great importance to pharmaceutical companies. This can be achieved by adding-on a mictrotracer within the existing early drug development programs. A microtracer quantity (around µg level) of 14C-labelled drug can be administered to subjects in any clinical trial. In order to quantify the drug and its related metabolites at these low levels an accelerator mass spectrometer (AMS) is used. Modern biomedical AMS facilities allow the analysis of more than 100 samples/day, down to low fg/mL levels. One of the upcoming applications of this technology is the generation of human metabolite profiles. According to the current guidelines, human metabolites can raise safety concerns when they are present at more than 10% of parent drug systemic exposure at steady state. Hence, knowledge on the number, quantity and identity of the formed metabolites for a given drug is of great importance. Traditionally, a dedicated clinical trial is required, as a consequence of the high radioactive dosing regimes, to obtain this information in a mass balance study. However, with modern AMS, any Phase I or II study can simultaneously become a metabolite in safety testing (MIST) study, simply by including a microtracer in the drug dose. True human data can be obtained, even before animal metabolic pathways have been established. Knowledge on the entire human metabolite profiles early in development, prevents surprises at later stages and will direct (pre)clinical development and toxicological testing. Here we will demonstrate the powerful combination of UPLC-High Resolution Mass Spectrometry (HRMS) and AMS analysis. Directly after UPLC separation the flow is diverted, whereby part is collected for off-line AMS analysis, and the remainder is directed to the HRMS for compound identification. All collected fractions are quantified for 14C levels by AMS. The reconstructed radiochromatogram enables detection and quantification of metabolites, while the HRMS enables metabolite identification. The combined use of AMS and HRMS on the same sample within even the same chromatogram clearly eliminates metID uncertainties. Simultaneous profile`acquisition on AMS and HRMS is extremely fast, powerful for metID, and besides, considerably more efficient, and affordable than conducting these two activities at separate facilities.

Combined use of HRMS and combustion AMS for simultaneous metabolite quantification and identification

253rd National Meeting (2017)

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