Application of a Rapid and Integrated Analysis System (RIAS) as a High-Throughput Processing Tool for In Vitro ADME Samples by Liquid Chromatography/Tandem Mass Spectrometry

Abstract

Over the past decade, drug discovery programs have started to address the optimization of key ADME properties already at an early stage of the process. Hence, analytical chemists have been confronted with tremendously rising sample numbers and have had to develop methodologies accelerating quantitative liquid chromatography/tandem mass spectrometry (LC/MS/MS). This article focuses on the application of a generic and fully automated LC/MS/MS, named Rapid and Integrated Analysis System (RIAS), as a high-throughput platform for the rapid quantification of drug-like compounds in various in vitro ADME assays. Previous efforts were dedicated to the setup and feasibility study of a workflow-integrated platform combining a modified high-throughput liquid handling LC/MS/MS system controlled by a customized software interface and a customized data-processing and reporting tool. Herein the authors present an extension of this previously developed basic application to a broad set of ADME screening campaigns, covering CYP inhibition, Caco-2, and PAMPA assays. The platform is capable of switching automatically between various ADME assays, performs MS compound optimization if required, and provides a speed of 8 s from sample to sample, independently of the type of ADME assay. Quantification and peak review are adopted to the high-throughput environment and tested against a standard HPLC-ESI technology.

Assessment of CYP3A‐mediated drug–drug interaction potential for victim drugs using an in vivo rat model

“…Then 150 μl of supernatant was transferred to a 2 ml deep well plate containing 150 μl of a solution with 20 ng/ml α-hydroxymidazolam-D4 in 50/50 (v/v) MeOH/H 2 O as internal standard. Formation of hydroxymidazolam was measured as described by Luippold et al [10].…”

Approach to Improve Compound Recovery in a High-Throughput Caco-2 Permeability Assay Supported by Liquid Chromatography–Tandem Mass Spectrometry

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