Solid-phase microextraction of endogenous metabolites from intact tissue validated using a Biocrates standard reference method kit
作者机构：Department of ChemistryUniversity of WaterlooWaterlooN2L 3G1Canada Department of Pharmacodynamics and Molecular PharmacologyFaculty of PharmacyCollegium Medicum in BydgoszczNicolaus Copernicus University in TorunBydgoszcz85-089Poland
出 版 物：《Journal of Pharmaceutical Analysis》 (药物分析学报（英文版）)
年 卷 期：2023年第13卷第1期
页 面：55-62页
核心收录：
学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 100704[医学-药物分析学] 1002[医学-临床医学] 0817[工学-化学工程与技术] 0703[理学-化学] 0702[理学-物理学] 10[医学]
基 金：supported by the Natural Sciences and Engineering Research Council of Canada NSERC(Grant No.:IRCPJ 184412-15)
主 题：Solid-phase microextraction Solvent extraction Metabolomics Sample preparation In vivo sampling
摘 要：Improved analytical methods for the metabolomic profiling of tissue samples are constantly ***,conventional sample preparation methods often involve tissue biopsy and/or homogenization,which disrupts the endogenous *** this study,solid-phase microextraction(SPME)fibers were used to monitor changes in endogenous compounds in homogenized and intact ovine lung *** SPME,a Biocrates AbsoluteIDQ assay was applied to make a downstream targeted metabolomics analysis and confirm the advantages of in vivo SPME *** AbsoluteIDQ kit enabled the targeted analysis of over 100 metabolites via solid-liquid extraction and *** analysis revealed significant differences between conventional liquid extractions from homogenized tissue and SPME results for both homogenized and intact tissue *** addition,principal component analysis revealed separated clustering among all the three sample groups,indicating changes in the metabolome due to tissue homogenization and the chosen sample preparation ***,clear differences in free metabolites were observed when extractions were performed on the intact and homogenized tissue using identical SPME ***,a direct comparison showed that 47 statistically distinct metabolites were detected between the homogenized and intact lung tissue samples(P0.05)using mixed-mode SPME *** changes were probably due to the disruptive homogenization of the *** study s findings highlight both the importance of sample preparation in tissue-based metabolomics studies and SPME s unique ability to perform minimally invasive extractions without tissue biopsy or homogenization while providing broad metabolite coverage.
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