{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,11]],"date-time":"2026-05-11T15:27:27Z","timestamp":1778513247457,"version":"3.51.4"},"reference-count":44,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2021,8,21]],"date-time":"2021-08-21T00:00:00Z","timestamp":1629504000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004281","name":"Narodowe Centrum Nauki","doi-asserted-by":"publisher","award":["Preludium no 2018\/29\/N\/NZ7\/02908"],"award-info":[{"award-number":["Preludium no 2018\/29\/N\/NZ7\/02908"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Metabolites"],"abstract":"<jats:p>Gastrointestinal stromal tumour has already been well explored at the genome level; however, little is known about metabolic processes occurring in the sarcoma. Sample preparation is a crucial step in untargeted metabolomics workflow, highly affecting the metabolome coverage and the quality of the results. In this study, four liquid-liquid extraction methods for the isolation of endogenous compounds from gastrointestinal stromal tumours were compared and evaluated. The protocols covered two-step or stepwise extraction with methyl-tert-butyl ether (MTBE) or dichloromethane. The extracts were subjected to LC-MS analysis by the application of reversed-phase and hydrophilic interaction liquid chromatography to enable the separation and detection of both polar and nonpolar analytes. The extraction methods were compared in terms of efficiency (total number of detected metabolites) and reproducibility. The method was based on the stepwise extraction with MTBE, methanol, and water proved to be the most reproducible, and thus, its robustness to fluctuations in experimental conditions was assessed employing Plackett\u2013Burman design and hierarchical modelling. While most studied factors had no effect on the metabolite abundance, the highest coefficient value was observed for the volume of MTBE added during extraction. Herein, we demonstrate the application and the feasibility of the selected protocol for the analysis of gastrointestinal stromal tumour samples. The method selected could be considered as a reference for the best characterization of underlying molecular changes associated with complex tissue extracts of GIST.<\/jats:p>","DOI":"10.3390\/metabo11080554","type":"journal-article","created":{"date-parts":[[2021,8,22]],"date-time":"2021-08-22T21:47:52Z","timestamp":1629668872000},"page":"554","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["A Robust Method for Sample Preparation of Gastrointestinal Stromal Tumour for LC\/MS Untargeted Metabolomics"],"prefix":"10.3390","volume":"11","author":[{"given":"Szymon","family":"Macioszek","sequence":"first","affiliation":[{"name":"Department of Biopharmaceutics and Pharmacodynamics, Medical University of Gda\u0144sk, Hallera 107, 80-416 Gda\u0144sk, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Danuta","family":"Dudzik","sequence":"additional","affiliation":[{"name":"Department of Biopharmaceutics and Pharmacodynamics, Medical University of Gda\u0144sk, Hallera 107, 80-416 Gda\u0144sk, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julia","family":"Jacyna","sequence":"additional","affiliation":[{"name":"Department of Biopharmaceutics and Pharmacodynamics, Medical University of Gda\u0144sk, Hallera 107, 80-416 Gda\u0144sk, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Agnieszka","family":"Wozniak","sequence":"additional","affiliation":[{"name":"Department of General Medical Oncology, Leuven Cancer Institute, University Hospitals Leuven, 3000 Leuven, Belgium"},{"name":"Laboratory of Experimental Oncology, Department of Oncology, KU Leuven, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5980-030X","authenticated-orcid":false,"given":"Patrick","family":"Sch\u00f6ffski","sequence":"additional","affiliation":[{"name":"Department of General Medical Oncology, Leuven Cancer Institute, University Hospitals Leuven, 3000 Leuven, Belgium"},{"name":"Laboratory of Experimental Oncology, Department of Oncology, KU Leuven, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1166-4817","authenticated-orcid":false,"given":"Micha\u0142 J.","family":"Markuszewski","sequence":"additional","affiliation":[{"name":"Department of Biopharmaceutics and Pharmacodynamics, Medical University of Gda\u0144sk, Hallera 107, 80-416 Gda\u0144sk, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,8,21]]},"reference":[{"key":"ref_1","first-page":"2740","article-title":"Metabolomics Applications in Precision Medicine: An Oncological Perspective","volume":"17","year":"2017","journal-title":"Curr. 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