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Application of Passive Sampling to Characterise the Fish Exometabolome

机译:被动采样在鱼类鱼类代谢组学研究中的应用

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摘要

The endogenous metabolites excreted by organisms into their surrounding environment, termed the exometabolome, are important for many processes including chemical communication. In fish biology, such metabolites are also known to be informative markers of physiological status. While metabolomics is increasingly used to investigate the endogenous biochemistry of organisms, no non-targeted studies of the metabolic complexity of fish exometabolomes have been reported to date. In environmental chemistry, Chemcatcher® (Portsmouth, UK) passive samplers have been developed to sample for micro-pollutants in water. Given the importance of the fish exometabolome, we sought to evaluate the capability of Chemcatcher® samplers to capture a broad spectrum of endogenous metabolites excreted by fish and to measure these using non-targeted direct infusion mass spectrometry metabolomics. The capabilities of C18 and styrene divinylbenzene reversed-phase sulfonated (SDB-RPS) Empore™ disks for capturing non-polar and polar metabolites, respectively, were compared. Furthermore, we investigated real, complex metabolite mixtures excreted from two model fish species, rainbow trout (Oncorhynchus mykiss) and three-spined stickleback (Gasterosteus aculeatus). In total, 344 biological samples and 28 QC samples were analysed, revealing 646 and 215 m/z peaks from trout and stickleback, respectively. The measured exometabolomes were principally affected by the type of Empore™ (Hemel Hempstead, UK) disk and also by the sampling time. Many peaks were putatively annotated, including several bile acids (e.g., chenodeoxycholate, taurocholate, glycocholate, glycolithocholate, glycochenodeoxycholate, glycodeoxycholate). Collectively these observations show the ability of Chemcatcher® passive samplers to capture endogenous metabolites excreted from fish.
机译:被生物体排泄到周围环境中的内源性代谢物,称为外代谢体,对于包括化学交流在内的许多过程都很重要。在鱼类生物学中,这类代谢物也是生理状态的信息标记。尽管代谢组学越来越多地用于研究生物体的内源性生物化学,但​​迄今为止,尚未有关于鱼类外生代谢组代谢复杂性的非针对性研究的报道。在环境化学领域,已经开发了Chemcatcher ®(英国朴次茅斯)无源采样器,用于采样水中的微量污染物。考虑到鱼类代谢组学的重要性,我们试图评估Chemcatcher ®采样器捕获鱼类排泄的各种内生代谢物并使用非靶向直接输注质谱代谢组学进行测量的能力。比较了C18和苯乙烯二乙烯基苯反相磺化(SDB-RPS)Empore™圆盘分别捕获非极性和极性代谢物的能力。此外,我们研究了从两种鳟鱼(虹鳟鱼(Oncorhynchus mykiss)和三棘棘鱼(Gasterosteus aculeatus))中排出的真实,复杂的代谢物混合物。总共分析了344个生物样品和28个QC样品,分别从鳟鱼和棘背鱼中发现了646和215 m / z峰。测得的表生代谢主要受Empore™(英国赫默尔亨普斯特德)圆片的类型以及采样时间的影响。推定注释了许多峰,包括几种胆汁酸(例如鹅去氧胆酸盐,牛磺胆酸盐,甘醇胆酸盐,甘草胆酸盐,糖基去氧胆酸盐,糖脱氧胆酸盐)。总的来说,这些观察结果表明Chemcatcher ®被动采样器能够捕获从鱼类排出的内源性代谢产物。

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