An optimised monophasic faecal extraction method for LC-MS analysis and its application in gastrointestinal disease
Kelly, Patricia E. and Jene Ng, H and Farrell, Gillian and McKirdy, Shona and Russell, Richard K. and Hansen, Richard and Rattray, Zahra and Gerasimidis, Konstantinos and Rattray, Nicholas J. W. (2022) An optimised monophasic faecal extraction method for LC-MS analysis and its application in gastrointestinal disease. Metabolites, 12 (11). 1110. ISSN 2218-1989 (https://doi.org/10.3390/metabo12111110)
Preview |
Text.
Filename: Kelly_etal_Metabolites_2022_An_optimised_monophasic_faecal_extraction_method_for_LC_MS_analysis.pdf
Final Published Version License: Download (1MB)| Preview |
Abstract
Liquid chromatography coupled with mass spectrometry (LC-MS) metabolomic approaches are widely used to investigate underlying pathogenesis of gastrointestinal disease and mechanism of action of treatments. However, there is an unmet requirement to assess faecal metabolite extraction methods for large-scale metabolomics studies. Current methods often rely on biphasic extractions using harmful halogenated solvents, making automation and large-scale studies challenging. The present study reports an optimised monophasic faecal extraction protocol that is suitable for untargeted and targeted LC-MS analyses. The impact of several experimental parameters, including sample weight, extraction solvent, cellular disruption method, and sample-to-solvent ratio, were investigated. It is suggested that a 50 mg freeze-dried faecal sample should be used in a methanol extraction (1:20) using bead beating as the means of cell disruption. This is revealed by a significant increase in number of metabolites detected, improved signal intensity, and wide metabolic coverage given by each of the above extraction parameters. Finally, we addressed the applicability of the method on faecal samples from patients with Crohn’s disease (CD) and coeliac disease (CoD), two distinct chronic gastrointestinal diseases involving metabolic perturbations. Untargeted and targeted metabolomic analysis demonstrated the ability of the developed method to detect and stratify metabolites extracted from patient groups and healthy controls (HC), highlighting characteristic changes in the faecal metabolome according to disease. The method developed is, therefore, suitable for the analysis of patients with gastrointestinal disease and can be used to detect and distinguish differences in the metabolomes of CD, CoD, and HC.
ORCID iDs
Kelly, Patricia E., Jene Ng, H, Farrell, Gillian, McKirdy, Shona, Russell, Richard K., Hansen, Richard, Rattray, Zahra ORCID: https://orcid.org/0000-0002-8371-8549, Gerasimidis, Konstantinos and Rattray, Nicholas J. W. ORCID: https://orcid.org/0000-0002-3528-6905;-
-
Item type: Article ID code: 83202 Dates: DateEvent14 November 2022Published14 November 2022Published Online7 November 2022AcceptedNotes: This article belongs to the Section Endocrinology and Clinical Metabolic Research Subjects: Medicine > Pharmacy and materia medica
Medicine > Internal medicine > Diseases of the digestive system. GastroenterologyDepartment: Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences Depositing user: Pure Administrator Date deposited: 15 Nov 2022 14:03 Last modified: 27 Sep 2024 01:25 Related URLs: URI: https://strathprints.strath.ac.uk/id/eprint/83202