An integrated platform for fully automated high-throughput LC–MS/MS analysis of in vitro metabolic stability assay samples

The evaluation of ADME properties contributes importantly to drug candidate selection and therefore is crucial for the drug discovery process. Providing a sufficiently high-throughput capability in laboratories dedicated to early pharmacokinetic studies will thereby shorten the entire drug discovery process. In this paper an integrated and fully automated LC–MS/MS-based platform is described, which enables the assessment of in vitro metabolic stability, a key ADME parameter. An ultra-rapid injection system was coupled to a triple quadrupole mass spectrometer and hard- and software were customized to perform sample identification, cleanup, MS compound optimization and sample measurement without manual interaction. Conventional chromatography was initially evaluated but was later replaced by solid phase extraction as the only purification step. Ultra-fast robotics, combined with a generic step gradient enables analysis times of 8 s/sample. Reproducibility and quality of data has been found fully comparable to data generated by validated LC–MS/MS methods. The system handles data for mass spectrometric compound optimization and MRM (Multiple Reaction Monitoring) analytics in a fully automated way and exhibits great potential for a generalized use in ADME replacing currently applied conventional LC–MS/MS approaches.

Graphical abstractA new system was setup and validated for the high-throughput LC–MS/MS analysis of metabolic stability samples in a real-life application with a speed of 8 s/sample (see figure for correlation validated LC–MS/MS system (x-axis) to new system (y-axis)).Figure optionsDownload full-size imageDownload high-quality image (90 K)Download as PowerPoint slide