Comparison of modified Matyash method to conventional solvent systems for polar metabolite and lipid extractions
Comparison of modified Matyash method to conventional solvent systems for polar metabolite and lipid extractions
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DOI：
10.1016/j.aca.2018.03.019
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发表时间：
2018-12-11
影响因子：
6.2
通讯作者：
Viant, Mark R.
中科院分区：
文献类型：
--
作者：
Sostare, Jelena;Di Guida, Riccardo;Viant, Mark R.
在过去的十年中，代谢组学在分析方法的通量和稳健性方面取得了重大进展。然而，生物流体和低质量组织样品的制备仍然是一个费力且可能不一致的手动过程，并且是高通量代谢组学的一个重要瓶颈。为了解决这一问题，我们比较了三种不同样品类型中的三种不同样品提取溶剂系统，目的是为随后的样品制备自动化选择最佳方案。我们研究并重新优化了最近引入的甲基叔丁基醚（MTBE）/甲醇/水溶剂体系中的溶剂比（这里称为修改的Matyash; 2.6/2.0/2.4，v/v/v），并将其与原始Matyash方法进行比较（10/3/2.5，v/v/v）和常规氯仿/甲醇/水（逐步Bligh和Dyer，2.0/2.0/1.8，v/v/v），使用两种生物流体（人血清和尿）和一种组织（完整的大型蚤）。这是首次报道使用Matyash方法从美国国立卫生研究院（NIH）模式生物D中提取代谢产物。magna.通过非靶向直接输注质谱代谢组学或LC-MS代谢组学分析提取的样品。总体而言，与原始Matyash方法（多1-29%的峰和多1-30%的代谢物）和Bligh和Dyer方法（多4-20%的峰和多1-41%的代谢物）相比，改良Matyash方法产生了更多的峰和purified注释的代谢物。此外，当考虑代谢物强度时，改良的Matyash方法具有上级优势。基于峰强度的mRSD较低的观察结果，改良Matyash方法的重现性高于其他方法（与原始Matyash方法相比，12个数据集中有10个数据集;与Bligh和Dyer方法相比，12个数据集中有8个数据集）。总之，与两种广泛使用的方法相比，改良的Matyash方法倾向于为本研究中的大多数样品类型提供更高的产率和重现性。(C)2018作者由爱思唯尔公司出版
In the last decade, metabolomics has experienced significant advances in the throughput and robustness of analytical methodologies. Yet the preparation of biofluids and low-mass tissue samples remains a laborious and potentially inconsistent manual process, and a significant bottleneck for high-throughput metabolomics. To address this, we have compared three different sample extraction solvent systems in three diverse sample types with the purpose of selecting an optimum protocol for subsequent automation of sample preparation. We have investigated and re-optimised the solvent ratios in the recently introduced methyl tert-butyl ether (MTBE)/methanol/water solvent system (here termed modified Matyash; 2.6/2.0/2.4, v/v/v) and compared it to the original Matyash method (10/3/2.5, v/v/v) and the conventional chloroform/methanol/water (stepwise Bligh and Dyer, 2.0/2.0/1.8, v/v/v) using two biofluids (human serum and urine) and one tissue (whole Daphnia magna). This is the first report of the use of the Matyash method for extracting metabolites from the US National Institutes of Health (NIH) model organism D. magna. Extracted samples were analysed by non-targeted direct infusion mass spectrometry metabolomics or LC-MS metabolomics. Overall, the modified Matyash method yielded a higher number of peaks and putatively annotated metabolites compared to the original Matyash method (1-29% more peaks and 1-30% more metabolites) and the Bligh and Dyer method (4-20% more peaks and 1-41% more metabolites). Additionally the modified Matyash method was superior when considering metabolite intensities. The reproducibility of the modified Matyash method was higher than other methods (in 10 out of 12 datasets, compared to the original Matyash method; and in 8 out of 12 datasets, compared to the Bligh and Dyer method), based upon the observation of a lower mRSD of peak intensities. In conclusion, the modified Matyash method tended to provide a higher yield and reproducibility for most sample types in this study compared to two widely used methods. (C) 2018 The Authors. Published by Elsevier B.V.