End-to-End High-Throughput Biotransformation Workflow: Automated Data Acquisition and Processing of Sub-Second UHPLC Peaks Using Multi-Reflecting Time-of-Flight Mass Spectrometry and Data-Mining
June 2026, ASMS Conference
Ismael Zamora1; Hania Khoury-Hollins2; Richard Lock2; David Pickles2; Robert S Plumb2; Ian Wilson3
1Mass Spec Analytica, Sant Cugat del Valles, Spain; 2Waters Corporation, Wilmslow, United Kingdom; 3Imperial College London, London, United Kingdom
Abstract
Introduction
Traditional ultra-high performance liquid chromatography (UHPLC™) paired with high-resolution mass spectrometry (HRMS) has long been the standard for drug metabolite characterization, but its throughput is limited by peak dispersion the need to balance mass resolution with scan speed, and data processing and rationalisation. These factors, along with labour-intensive manual data review, slow down the delivery of actionable results.
To overcome these bottlenecks, we introduce a fully integrated high-throughput data dependent workflow combining a next generation UHPLC with multi reflecting time-of-flight technology (Xevo™ MRT MS) and dedicated data-mining software. This platform enables rapid, automated acquisition and processing of sub-second UHPLC peaks, achieving part-per-billion mass accuracy (≤500 ppb RMS), scan speeds up to 100 Hz, and high mass resolution (100K FWHM).
Methods
Male beagle dogs received a single intravenous (IV) dose, and whole blood was collected from the jugular vein both before and after dosing. Plasma was obtained by centrifugation and analyzed using reversed-phase UHPLC coupled to multi reflecting time of flight mass spectrometer.
Data dependent acquisition mode was used, and metabolites were detected and quantified in both positive and negative electrospray ionization modes, utilizing Xevo MRT MS and a dedicated MassMetaSite software.
Workflow improvements were assessed by comparing results obtained with shorter runtimes to those from conventional UHPLC method used by the contract research organization (CRO) that prepared the samples. This comparison focused on data quality, sample analysis times, and confidence in structural characterization.
You must be logged in to access this content. Not yet registered? Create a new account