Development of a single mobile phase for LC-IM-MS-based discovery lipidomics and metabolic phenotyping: Application to methapyrilene hepatotoxicity in the rat.

Autor: Wilson ID; Computational & Systems Medicine, Department of Metabolism, Digestion and Reproduction, Imperial College, Burlington Danes Building, Du Cane Road, London, W12 0NN, UK., Broeckling C; Bioanalysis and Omics, Colorado State University, Fort Collins, CO 8052, USA., Gethings LA; Waters Corporation, Stamford Ave, Wilmslow, SK9 4AX, UK., Munjoma NC; Waters Corporation, Stamford Ave, Wilmslow, SK9 4AX, UK., Trengove R; Curtin Health Innovation Research Institute, Curtin University, Kent St, Bentley WA 6102, Australia. Electronic address: Robert.Trengove@curtin.edu.au., Rainville PD; Waters Corporation, Milford, MA 01757, USA., Lai SK; Waters Corporation, Milford, MA 01757, USA., Isaac G; Program in Molecular Medicine, University of Massachusetts Chan Medical School, Worcester, MA 01605, USA., Plumb RS; Waters Corporation, Milford, MA 01757, USA. Electronic address: Rob_Plumb@waters.com.
Jazyk: angličtina
Zdroj: Journal of chromatography. A [J Chromatogr A] 2024 Jan 11; Vol. 1714, pp. 464552. Date of Electronic Publication: 2023 Dec 10.
DOI: 10.1016/j.chroma.2023.464552
Abstrakt: The untargeted global profiling of endogenous metabolites and lipids has the potential to increase knowledge and understanding in many areas of biology. LC-MS/MS is a key technology for such analyses however, several different LC methodologies, using different mobile phase compositions, are required to cover the diversity in polarity and analyte structure encountered in biological samples. Most notably many lipid screening methods make use of isopropanol (IPA) as a major component of mobile phases employed for comprehensive lipidomic profiling. In order to increase laboratory efficiency, and minimize opportunities for errors, a suite of methods, based on a single acetonitrile (ACN)-aqueous buffer mobile phase combination, has been developed. This mobile phase can be used for hydrophobic interaction liquid chromatography on an amide stationary phase (for polar analytes), reversed-phase (RP) LC analysis on a C8 stationary phase (for moderately polar-non-polar compounds) and RPLC using a CSH phenyl-hexyl bonded column (for lipids). All of these sub 10 minute separations had good throughput and reproducibility with CV's of analyte response <25 % whilst eliminating the need for complex mobile phase preparation and the use of IPA as an organic modifier for lipidomics. Advantages of removing IPA and replacing it with the ACN-based method were a 58 % increase in peak capacity for lipids, with improved resolution for the di- and triglycerides and cholesterol esters compared to current methods. Compared to the IPA-containing solvent system the ACN-based mobile phase also resulted in a 61 % increase in lipid feature detection. The utility of this "universal" mobile phase approach was demonstrated by its application to a rat toxicology study investigating the consequences of methapyrilene administration through on the endogenous metabolite profiles of plasma and urine. Methapyrilene and its metabolites were also profiled in these samples.
Competing Interests: Declaration of Competing Interest IW acts as a consultant for a number of organisations including Waters Corporation. The remaining authors declare no conflicts of interest.
(Copyright © 2023. Published by Elsevier B.V.)
Databáze: MEDLINE
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