Procedures for large-scale metabolic profiling of serum and plasma using gas chromatography and liquid chromatography coupled to mass spectrometry

被引：2910

作者：

Dunn, Warwick B. ^{[1
,2
,3
,4
]}

Broadhurst, David ^{[2
,5
]}

Begley, Paul ^{[2
]}

Zelena, Eva ^{[2
]}

Francis-McIntyre, Sue ^{[2
]}

Anderson, Nadine ^{[2
]}

Brown, Marie ^{[2
]}

Knowles, Joshau D. ^{[6
]}

Halsall, Antony ^{[2
]}

Haselden, John N. ^{[7
]}

Nicholls, Andrew W. ^{[7
]}

Wilson, Ian D. ^{[8
]}

Kell, Douglas B. ^{[2
]}

Goodacre, Royston ^{[1
,2
]}

机构：

[1] Univ Manchester, Manchester Interdisciplinary Bioctr, Manchester Ctr Integrat Syst Biol, Manchester, Lancs, England

[2] Univ Manchester, Manchester Interdisciplinary Bioctr, Sch Chem, Manchester, Lancs, England

[3] Manchester Biomed Res Ctr, Ctr Adv Discoveries & Expt Therapeut, Manchester, Lancs, England

[4] Sch Biomed, Manchester, Lancs, England

[5] Univ Alberta, Dept Med, Katz Grp Ctr Pharm & Hlth, Edmonton, AB, Canada

[6] Univ Manchester, Sch Comp Sci, Manchester, Lancs, England

[7] GlaxoSmithKline, Dept Invest Preclin Toxicol, London, Herts, England

[8] AstraZeneca, Dept Clin Pharmacol Drug Metab & Pharmacokinet, Macclesfield, Cheshire, England

来源：

NATURE PROTOCOLS | 2011年 / 6卷 / 07期

基金：

英国生物技术与生命科学研究理事会; 英国工程与自然科学研究理事会;

关键词：

SYSTEMS BIOLOGY; TOF-MS; HYDROPHILIC INTERACTION; COMPREHENSIVE ANALYSIS; HUMAN URINE; HPLC-MS; GC-TOF; PERFORMANCE; BIOMARKERS; DERIVATIZATION;

D O I：

10.1038/nprot.2011.335

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

Metabolism has an essential role in biological systems. Identification and quantitation of the compounds in the metabolome is defined as metabolic profiling, and it is applied to define metabolic changes related to genetic differences, environmental influences and disease or drug perturbations. Chromatography-mass spectrometry (MS) platforms are frequently used to provide the sensitive and reproducible detection of hundreds to thousands of metabolites in a single biofluid or tissue sample. Here we describe the experimental workflow for long-term and large-scale metabolomic studies involving thousands of human samples with data acquired for multiple analytical batches over many months and years. Protocols for serum-and plasma-based metabolic profiling applying gas chromatography-MS (GC-MS) and ultraperformance liquid chromatography-MS (UPLC-MS) are described. These include biofluid collection, sample preparation, data acquisition, data pre-processing and quality assurance. Methods for quality control-based robust LOESS signal correction to provide signal correction and integration of data from multiple analytical batches are also described.

引用

页码：1060 / 1083

页数：24

共 100 条

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High-throughput classification of yeast mutants for functional genomics using metabolic footprinting [J].