Comprehensive online multicolumn two-dimensional liquid chromatography-diode array detection-mass spectrometry workflow as a framework for chromatographic screening and analysis of new drug substances
- 4 March 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Analytical and Bioanalytical Chemistry
- Vol. 412 (11), 2655-2663
- https://doi.org/10.1007/s00216-020-02498-8
Abstract
The analysis of complex mixtures of closely related species is quickly becoming a bottleneck in the development of new drug substances, reflecting the ever-increasing complexity of both fundamental biology and the therapeutics used to treat disease. Two-dimensional liquid chromatography (2D-LC) is emerging as a powerful tool to achieve substantial improvements in peak capacity and selectivity. However, 2D-LC suffers from several limitations, including the lack of automated multicolumn setups capable of combining multiple columns in both dimensions. Herein, we report an investigation into the development and implementation of a customized online comprehensive multicolumn 2D-LC-DAD-MS setup for screening and method development purposes, as well as analysis of multicomponent biopharmaceutical mixtures. In this study, excellent chromatographic performance in terms of selectivity, peak shape, and reproducibility were achieved by combining reversed-phase (RP), strong cation exchange (SCX), strong anion exchange (SAX), and size exclusion chromatography (SEC) using sub-2-μm columns in the first dimension in conjunction with several 3.0 mm × 50 mm RP columns packed with sub-3-μm fully porous particles in the second dimension. Multiple combinations of separation modes coupled to UV and MS detection are applied to the LC × LC analysis of a protein standard mixture, intended to be representative of protein drug substances. The results reported in this study demonstrate that our automated online multicolumn 2D-LC-DAD-MS workflow can be a powerful tool for comprehensive chromatographic column screening that enables the semi-automated development of 2D-LC methods, offering the ability to streamline full visualization of sample composition for an unknown complex mixture while maximizing chromatographic orthogonality. Graphical AbstractKeywords
This publication has 21 references indexed in Scilit:
- Column selection for comprehensive two-dimensional liquid chromatography using the hydrophobic subtraction modelJournal of Chromatography A, 2019
- High Throughput Strategies for the Discovery and Optimization of Catalytic ReactionsAngewandte Chemie, 2018
- First inter-laboratory study of a Supercritical Fluid Chromatography method for the determination of pharmaceutical impuritiesJournal of Pharmaceutical and Biomedical Analysis, 2018
- Ligand libraries for high throughput screening of homogeneous catalystsChemical Society Reviews, 2018
- Visualizing and studying frictional heating effects in reversed-phase liquid chromatography using infrared thermal imagingAnalytica Chimica Acta, 2018
- Optimizing separations in online comprehensive two‐dimensional liquid chromatographyJournal of Separation Science, 2017
- Current challenges and future prospects in chromatographic method development for pharmaceutical researchTrAC Trends in Analytical Chemistry, 2017
- Two-Dimensional Liquid Chromatography: A State of the Art TutorialAnalytical Chemistry, 2016
- Ultrafast chiral separations for high throughput enantiopurity analysisChemical Communications, 2016
- Genotoxic Impurities in Pharmaceutical Manufacturing: Sources, Regulations, and MitigationChemical Reviews, 2015