title: Establishment of Protocols for Global Metabolomics by LC-MS for Biomarker Discovery.
abstract: 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.
authors: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Miyagi, Japan.|Medical Biochemistry, Tohoku University School of Medicine, Sendai, Miyagi, Japan.|CREST, Japan Agency for Medical Research and Development (AMED), Chiyoda, Tokyo, Japan.
authors: Daisuke
authors: http://orcid.org/0000-0001-9484-9870
authors: D
authors: Saigusa
authors: Department of Systems Bioinformatics, Graduate School of Information Sciences, Tohoku University, Sendai, Miyagi, Japan.
authors: Yasunobu
authors: Y
authors: Okamura
authors: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Miyagi, Japan.|Department of Systems Bioinformatics, Graduate School of Information Sciences, Tohoku University, Sendai, Miyagi, Japan.
authors: Ikuko N
authors: IN
authors: Motoike
authors: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Miyagi, Japan.|Department of Biochemistry, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.
authors: Yasutake
authors: Y
authors: Katoh
authors: Department of Gynecology and Obstetrics, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.
authors: Yasuhiro
authors: Y
authors: Kurosawa
authors: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Miyagi, Japan.
authors: Reina
authors: R
authors: Saijyo
authors: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Miyagi, Japan.|Medical Biochemistry, Tohoku University School of Medicine, Sendai, Miyagi, Japan.
authors: Seizo
authors: S
authors: Koshiba
authors: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Miyagi, Japan.
authors: Jun
authors: J
authors: Yasuda
authors: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Miyagi, Japan.|Department of Gene Expression Regulation, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Miyagi, Japan.
authors: Hozumi
authors: H
authors: Motohashi
authors: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Miyagi, Japan.|Department of Gynecology and Obstetrics, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.
authors: Junichi
authors: J
authors: Sugawara
authors: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Miyagi, Japan.
authors: Osamu
authors: O
authors: Tanabe
authors: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Miyagi, Japan.|Department of Systems Bioinformatics, Graduate School of Information Sciences, Tohoku University, Sendai, Miyagi, Japan.
authors: Kengo
authors: K
authors: Kinoshita
authors: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Miyagi, Japan.|Medical Biochemistry, Tohoku University School of Medicine, Sendai, Miyagi, Japan.
authors: Masayuki
authors: M
authors: Yamamoto
mesh_terms: D015415:Biomarkers / Q000097:blood; D002853:Chromatography, Liquid / Q000379:methods / Q000592:standards; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards*; D055432:Metabolomics / Q000379:methods* / Q000592:standards*
chemical_list: D015415:Biomarkers