Towards Standards for Human Fecal Sample Preparation in Targeted and Untargeted LC-HRMS Studies.
Towards Standards for Human Fecal Sample Preparation in Targeted and Untargeted LC-HRMS Studies.
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DOI：
10.3390/metabo11060364
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发表时间：
2021-06-07
期刊：
影响因子：
4.1
通讯作者：
Hankemeier T
中科院分区：
文献类型：
--
作者：
Hosseinkhani F;Dubbelman AC;Karu N;Harms AC;Hankemeier T
肠道微生物群及其代谢产物越来越被认为是人类健康的重要调节剂。粪便代谢组提供了健康和疾病中人体代谢与肠道微生物群之间相互作用的功能读数。由于粪便基质的高度复杂性，样品制备通常会引入技术变化，必须将其最小化以准确检测和定量肠道细菌代谢物。在这里，我们测试了六种不同的代表性浸提方法（单相和液-液浸提），并比较了粪便量、浸提溶剂类型和溶剂pH值的差异。我们的结果表明，需要最小粪便（湿）量为0.50 g才能准确表示粪便的复杂质地。结果表明，MTBE法（MTBE/甲醇/水，3.6/2.8/3.5，v/v/v）对11种不同类型的化合物具有最高的提取效率，提取的特征量最多（占不同提取物中总特征量的97%），重复性（CV < 35%）和提取回收率（≥70%）优于其他提取方法。重要的是，优化每个步骤的溶剂体积至初始干燥粪便物质（µL/mg粪便）是迈向标准化的重要一步，这使得能够自信地评估肠道细菌代谢物对人类健康的贡献。
Gut microbiota and their metabolic products are increasingly being recognized as important modulators of human health. The fecal metabolome provides a functional readout of the interactions between human metabolism and the gut microbiota in health and disease. Due to the high complexity of the fecal matrix, sample preparation often introduces technical variation, which must be minimized to accurately detect and quantify gut bacterial metabolites. Here, we tested six different representative extraction methods (single-phase and liquid–liquid extractions) and compared differences due to fecal amount, extraction solvent type and solvent pH. Our results indicate that a minimum fecal (wet) amount of 0.50 g is needed to accurately represent the complex texture of feces. The MTBE method (MTBE/methanol/water, 3.6/2.8/3.5, v/v/v) outperformed the other extraction methods, reflected by the highest extraction efficiency for 11 different classes of compounds, the highest number of extracted features (97% of the total identified features in different extracts), repeatability (CV < 35%) and extraction recovery (≥70%). Importantly, optimization of the solvent volume of each step to the initial dried fecal material (µL/mg feces) offers a major step towards standardization, which enables confident assessment of the contributions of gut bacterial metabolites to human health.
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影响因子：
12.2
作者：
Hosseinkhani F;Heinken A;Thiele I;Lindenburg PW;Harms AC;Hankemeier T
通讯作者：
Hankemeier T
影响因子：
3.6
作者：
Buijink, M. Renate;van Weeghel, Michel;Michel, Stephan
通讯作者：
Michel, Stephan
影响因子：
6.1
作者：
Lopez-Bascon, M. A.;Calderon-Santiago, M.;Priego-Capote, F.
通讯作者：
Priego-Capote, F.
影响因子：
6.2
作者：
Sostare, Jelena;Di Guida, Riccardo;Viant, Mark R.
通讯作者：
Viant, Mark R.
影响因子：
4.6
作者：
Nagpal R;Wang S;Ahmadi S;Hayes J;Gagliano J;Subashchandrabose S;Kitzman DW;Becton T;Read R;Yadav H
通讯作者：
Yadav H