Establishment of Protocols for Global Metabolomics by LC-MS for Biomarker Discovery.
Establishment of Protocols for Global Metabolomics by LC-MS for Biomarker Discovery.
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DOI：
10.1371/journal.pone.0160555
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Yamamoto M
中科院分区：
文献类型：
--
作者：
Saigusa D;Okamura Y;Motoike IN;Katoh Y;Kurosawa Y;Saijyo R;Koshiba S;Yasuda J;Motohashi H;Sugawara J;Tanabe O;Kinoshita K;Yamamoto M
代谢组学是一种很有前途的生物标志物发现途径。尽管代谢组学分析的质量，特别是使用质谱（MS）的全球代谢组学（G-Met）的质量在很大程度上取决于仪器，但代谢组学工作流程中的几个基本层面仍然存在潜在的瓶颈。因此，为了克服这些困难，我们初步建立了一套精确的人体血浆G-Met分析方案。在我们的方案中，样品在96孔板中使用自动液体处理系统进行脱蛋白，并使用配备反相柱的UHPLC-QTOF/MS系统或配备正相柱的LC-FTMS系统进行。我们还制定了G-Met数据的归一化方案，以补偿批内和批间的差异，随着我们的归一化，差异显著减少，特别是对于使用C18反相柱的UHPLC-QTOF/MS数据。其次，我们检测了edta血液标本储存引起的代谢组学变化，以确定用于评估标本分析前条件的质量标记。采用G-Met法鉴定溶血磷脂、二肽、脂肪酸、琥珀酸、氨基酸、葡萄糖、尿酸等40种质量标记物，评价血浆样品质量，建立质量评分计算公式。我们将我们的质量标记应用于一项小规模研究，以评估临床样本的质量。这里建立的G-Met协议和质量标记应该证明对更广泛疾病的生物标记物的发现和开发是有用的。
Metabolomics is a promising avenue for biomarker discovery. Although the quality of metabolomic analyses, especially global metabolomics (G-Met) using mass spectrometry (MS), largely depends on the instrumentation, potential bottlenecks still exist at several basic levels in the metabolomics workflow. Therefore, we established a precise protocol initially for the G-Met analyses of human blood plasma to overcome some these difficulties. In our protocol, samples are deproteinized in a 96-well plate using an automated liquid-handling system, and conducted either using a UHPLC-QTOF/MS system equipped with a reverse phase column or a LC-FTMS system equipped with a normal phase column. A normalization protocol of G-Met data was also developed to compensate for intra- and inter-batch differences, and the variations were significantly reduced along with our normalization, especially for the UHPLC-QTOF/MS data with a C18 reverse-phase column for positive ions. Secondly, we examined the changes in metabolomic profiles caused by the storage of EDTA-blood specimens to identify quality markers for the evaluation of the specimens’ pre-analytical conditions. Forty quality markers, including lysophospholipids, dipeptides, fatty acids, succinic acid, amino acids, glucose, and uric acid were identified by G-Met for the evaluation of plasma sample quality and established the equation of calculating the quality score. We applied our quality markers to a small-scale study to evaluate the quality of clinical samples. The G-Met protocols and quality markers established here should prove useful for the discovery and development of biomarkers for a wider range of diseases.
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影响因子：
30.8
作者：
Kettunen, Johannes;Tukiainen, Taru;Sarin, Antti-Pekka;Ortega-Alonso, Alfredo;Tikkanen, Emmi;Lyytikainen, Leo-Pekka;Kangas, Antti J.;Soininen, Pasi;Wuertz, Peter;Silander, Kaisa;Dick, Danielle M.;Rose, Richard J.;Savolainen, Markku J.;Viikari, Jorma;Kahonen, Mika;Lehtimaki, Terho;Pietilainen, Kirsi H.;Inouye, Michael;McCarthy, Mark I.;Jula, Antti;Eriksson, Johan;Raitakari, Olli T.;Salomaa, Veikko;Kaprio, Jaakko;Jarvelin, Marjo-Riitta;Peltonen, Leena;Perola, Markus;Freimer, Nelson B.;Ala-Korpela, Mika;Palotie, Aarno;Ripatti, Samuli
通讯作者：
Ripatti, Samuli
影响因子：
30.8
作者：
Shin, So-Youn;Fauman, Eric B.;Petersen, Ann-Kristin;Krumsiek, Jan;Santos, Rita;Huang, Jie;Arnold, Matthias;Erte, Idil;Forgetta, Vincenzo;Yang, Tsun-Po;Walter, Klaudia;Menni, Cristina;Chen, Lu;Vasquez, Louella;Valdes, Ana M.;Hyde, Craig L.;Wang, Vicky;Ziemek, Daniel;Roberts, Phoebe;Xi, Li;Grundberg, Elin;Waldenberger, Melanie;Richards, J. Brent;Mohney, Robert P.;Milburn, Michael V.;John, Sally L.;Trimmer, Jeff;Theis, Fabian J.;Overington, John P.;Suhre, Karsten;Brosnan, M. Julia;Gieger, Christian;Kastenmueller, Gabi;Spector, Tim D.;Soranzo, Nicole
通讯作者：
Soranzo, Nicole
影响因子：
3.6
作者：
Martin, Jean-Charles;Maillot, Matthieu;Mazerolles, Gerard;Verdu, Alexandre;Lyan, Bernard;Migne, Carole;Defoort, Catherine;Canlet, Cecile;Junot, Christophe;Guillou, Claude;Manach, Claudine;Jabob, Daniel;Bouveresse, Delphine Jouan-Rimbaud;Paris, Estelle;Pujos-Guillot, Estelle;Jourdan, Fabien;Giacomoni, Franck;Courant, Frederique;Fave, Gaelle;Le Gall, Gwenaelle;Chassaigne, Hubert;Tabet, Jean-Claude;Martin, Jean-Francois;Antignac, Jean-Philippe;Shintu, Laetitia;Defernez, Marianne;Philo, Mark;Alexandre-Gouaubau, Marie-Cecile;Amiot-Carlin, Marie-Josephe;Bossis, Mathilde;Triba, Mohamed N.;Stojilkovic, Natali;Banzet, Nathalie;Molinie, Roland;Bott, Romain;Goulitquer, Sophie;Caldarelli, Stefano;Rutledge, Douglas N.
通讯作者：
Rutledge, Douglas N.
影响因子：
4.1
作者：
Stanberry L;Mias GI;Haynes W;Higdon R;Snyder M;Kolker E
通讯作者：
Kolker E
影响因子：
3
作者：
Kind T;Fiehn O
通讯作者：
Fiehn O