Addressing the analytical throughput challenges in ADME screening using rapid ultra‐performance liquid chromatography/tandem mass spectrometry methodologies

Abstract

High-throughput ADME screening for compound drug development properties has become an essential part of the modern drug discovery process, allowing more informed decisions to be made on the best compounds to take forward in the discovery/development process. This however is a time-consuming process requiring multiple tests to be performed, demanding a significant amount of liquid chromatography/mass spectrometry (LC/MS) instrument time. This article focuses on the use of sub-2 microm porous particle LC coupled to tandem quadrupole MS/MS mass spectrometry for the rapid screening of ADME properties. Using this approach analysis times from 30 s to 1 min were achievable allowing analysis times to be cut by 80%. The use of the small particles coupled to high flow rates allowed for sufficient resolution, even with very short analysis time, to resolve the analytes of interest from similar compounds that would interfere with the assay. The use of dedicated, intelligent, software packages allowed for the user-free generation of MS/MS conditions and the processing of the data.

Comparison of liquid chromatography and supercritical fluid chromatography coupled to compact single quadrupole mass spectrometer for targeted in vitro metabolism assay

“…Among the untargeted studies, metabolite profiling and metabolite identification (metabolite ID) have to be carried out with high-end MS devices (i.e., orbitrap or QqTOF/MS) for investigating the numerous unknown metabolites derived from metabolism [8,9]. Over the past few years, significant efforts have been made to improve the acquisition rate, sensitivity and resolving power of high-resolution MS instruments, which are generally employed for these more sophisticated in vitro metabolism studies [10][11][12]. The most recent developments of quadrupole-based instruments have been focused on making them more sensitive and user-friendly, and the most widely used strategy for simple in vitro metabolism assay consists in performing generic LC conditions combined with state-of the-art MS/MS detector, to achieve a sufficient level of sensitivity and selectivity.…”

LC‐MS in Drug Metabolism and Pharmacokinetics: A Pharmaceutical Industry Perspective

“…Bioanalytical scientists that support ADME screens are constantly searching for ways to improve LC -MS analysis speed. High -throughput LC -MS bioanalytical approaches such as high speed separation on short columns packed with small particles [52] , ultrahigh pressure liquid chromatography ( UHPLC ) using sub -2 μ m [53,54] or other high effi ciency columns such as fused core columns [55] , are all very much applicable in ADME This process of obtaining an optimal SRM transition and corresponding lens voltages is a prerequisite for achieving the best sensitivity and specifi city for quantitative bioanalysis, and has to be performed for each individual test compound of interest. Usually, SRM optimization is conducted manually by infusing a compound solution into the instrument while the mass spectrometer is scanning, and ramping individual parameters to obtain their optimum values, at a throughput of about 5 -10 min per compound.…”

“…In ADME bioanalysis, on the other hand, several dozens of compounds are commonly assayed and analyzed within a single batch, and as a result, the serial workfl ow designed for single -compound analysis is not adequate. Over the years, vendors have developed high -throughput software tools to manage the ADME workfl ow and enable batch mode MS/MS optimization, sample analysis, and data reviewing [54] . Previously mentioned software such as QuickQuan [49] and DiscoveryQuant [51] not only provides automated optimization for SRM MS/MS methods, but also is capable of storing the optimized conditions in a shared database, generating acquisition and processing methods, performing sample analysis, and processing results in an automated fashion.…”

Current Technology and Limitation of LC‐MS

“…The ultimate advantage is increased chromatographic effi ciency and shorter cycle times and the latter will increase the capacity of the LC -MS instrument (Mazzeo et al, 2005 ;Plumb et al, 2008 ). Moreover, short gradients may come at the expense of chromatographic resolution, which could be problematic for identifi cation of isobaric metabolites (e.g., if multiple hydroxylated metabolites are formed).…”