Optimized Analytical Procedures for the Untargeted Metabolomic Profiling of Human Urine and Plasma by Combining Hydrophilic Interaction (HILIC) and Reverse-Phase Liquid Chromatography (RPLC)-Mass Spectrometry

Overview

Journal Mol Cell Proteomics

Specialties Biochemistry
Cell Biology
Molecular Biology

Date 2015 Mar 20

PMID 25787789

Citations 78

Authors

Kevin Contrepois

Lihua Jiang

Michael Snyder

Affiliations

Soon will be listed here.

Abstract

Profiling of body fluids is crucial for monitoring and discovering metabolic markers of health and disease and for providing insights into human physiology. Since human urine and plasma each contain an extreme diversity of metabolites, a single liquid chromatographic system when coupled to mass spectrometry (MS) is not sufficient to achieve reasonable metabolome coverage. Hydrophilic interaction liquid chromatography (HILIC) offers complementary information to reverse-phase liquid chromatography (RPLC) by retaining polar metabolites. With the objective of finding the optimal combined chromatographic solution to profile urine and plasma, we systematically investigated the performance of five HILIC columns with different chemistries operated at three different pH (acidic, neutral, basic) and five C18-silica RPLC columns. The zwitterionic column ZIC-HILIC operated at neutral pH provided optimal performance on a large set of hydrophilic metabolites. The RPLC columns Hypersil GOLD and Zorbax SB aq were proven to be best suited for the metabolic profiling of urine and plasma, respectively. Importantly, the optimized HILIC-MS method showed excellent intrabatch peak area reproducibility (CV < 12%) and good long-term interbatch (40 days) peak area reproducibility (CV < 22%) that were similar to those of RPLC-MS procedures. Finally, combining the optimal HILIC- and RPLC-MS approaches greatly expanded metabolome coverage with 44% and 108% new metabolic features detected compared with RPLC-MS alone for urine and plasma, respectively. The proposed combined LC-MS approaches improve the comprehensiveness of global metabolic profiling of body fluids and thus are valuable for monitoring and discovering metabolic changes associated with health and disease in clinical research studies.

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References

Cubbon S, Bradbury T, Wilson J, Thomas-Oates J . Hydrophilic interaction chromatography for mass spectrometric metabonomic studies of urine. Anal Chem. 2007; 79(23):8911-8. DOI: 10.1021/ac071008v. View

Nordstrom A, Want E, Northen T, Lehtio J, Siuzdak G . Multiple ionization mass spectrometry strategy used to reveal the complexity of metabolomics. Anal Chem. 2007; 80(2):421-9. DOI: 10.1021/ac701982e. View

Sreekumar A, Poisson L, Rajendiran T, Khan A, Cao Q, Yu J . Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression. Nature. 2009; 457(7231):910-4. PMC: 2724746. DOI: 10.1038/nature07762. View

Fujieda Y, Manno A, Hayashi Y, Rhodes N, Guo L, Arita M . Inflammation and resolution are associated with upregulation of fatty acid β-oxidation in Zymosan-induced peritonitis. PLoS One. 2013; 8(6):e66270. PMC: 3679047. DOI: 10.1371/journal.pone.0066270. View

Want E, Wilson I, Gika H, Theodoridis G, Plumb R, Shockcor J . Global metabolic profiling procedures for urine using UPLC-MS. Nat Protoc. 2010; 5(6):1005-18. DOI: 10.1038/nprot.2010.50. View

Psychogios N, Hau D, Peng J, Guo A, Mandal R, Bouatra S . The human serum metabolome. PLoS One. 2011; 6(2):e16957. PMC: 3040193. DOI: 10.1371/journal.pone.0016957. View

Kaddurah-Daouk R, Kristal B, Weinshilboum R . Metabolomics: a global biochemical approach to drug response and disease. Annu Rev Pharmacol Toxicol. 2008; 48:653-83. DOI: 10.1146/annurev.pharmtox.48.113006.094715. View

Spagou K, Tsoukali H, Raikos N, Gika H, Wilson I, Theodoridis G . Hydrophilic interaction chromatography coupled to MS for metabonomic/metabolomic studies. J Sep Sci. 2010; 33(6-7):716-27. DOI: 10.1002/jssc.200900803. View

Patti G, Yanes O, Siuzdak G . Innovation: Metabolomics: the apogee of the omics trilogy. Nat Rev Mol Cell Biol. 2012; 13(4):263-9. PMC: 3682684. DOI: 10.1038/nrm3314. View

10.

Buscher J, Czernik D, Ewald J, Sauer U, Zamboni N . Cross-platform comparison of methods for quantitative metabolomics of primary metabolism. Anal Chem. 2009; 81(6):2135-43. DOI: 10.1021/ac8022857. View

11.

Pujos-Guillot E, Hubert J, Martin J, Lyan B, Quintana M, Claude S . Mass spectrometry-based metabolomics for the discovery of biomarkers of fruit and vegetable intake: citrus fruit as a case study. J Proteome Res. 2013; 12(4):1645-59. DOI: 10.1021/pr300997c. View

12.

Kuehnbaum N, Britz-McKibbin P . New advances in separation science for metabolomics: resolving chemical diversity in a post-genomic era. Chem Rev. 2013; 113(4):2437-68. DOI: 10.1021/cr300484s. View

13.

Dunn W, Broadhurst D, Begley P, Zelena E, Francis-McIntyre S, Anderson N . Procedures for large-scale metabolic profiling of serum and plasma using gas chromatography and liquid chromatography coupled to mass spectrometry. Nat Protoc. 2011; 6(7):1060-83. DOI: 10.1038/nprot.2011.335. View

14.

Townsend M, Clish C, Kraft P, Wu C, Souza A, Deik A . Reproducibility of metabolomic profiles among men and women in 2 large cohort studies. Clin Chem. 2013; 59(11):1657-67. PMC: 3812240. DOI: 10.1373/clinchem.2012.199133. View

15.

Zhang A, Sun H, Han Y, Yan G, Yuan Y, Song G . Ultraperformance liquid chromatography-mass spectrometry based comprehensive metabolomics combined with pattern recognition and network analysis methods for characterization of metabolites and metabolic pathways from biological data sets. Anal Chem. 2013; 85(15):7606-12. DOI: 10.1021/ac401793d. View

16.

Roux A, Xu Y, Heilier J, Olivier M, Ezan E, Tabet J . Annotation of the human adult urinary metabolome and metabolite identification using ultra high performance liquid chromatography coupled to a linear quadrupole ion trap-Orbitrap mass spectrometer. Anal Chem. 2012; 84(15):6429-37. DOI: 10.1021/ac300829f. View

17.

Smith C, Want E, OMaille G, Abagyan R, Siuzdak G . XCMS: processing mass spectrometry data for metabolite profiling using nonlinear peak alignment, matching, and identification. Anal Chem. 2006; 78(3):779-87. DOI: 10.1021/ac051437y. View

18.

Spagou K, Wilson I, Masson P, Theodoridis G, Raikos N, Coen M . HILIC-UPLC-MS for exploratory urinary metabolic profiling in toxicological studies. Anal Chem. 2010; 83(1):382-90. DOI: 10.1021/ac102523q. View

19.

Yuan M, Breitkopf S, Yang X, Asara J . A positive/negative ion-switching, targeted mass spectrometry-based metabolomics platform for bodily fluids, cells, and fresh and fixed tissue. Nat Protoc. 2012; 7(5):872-81. PMC: 3685491. DOI: 10.1038/nprot.2012.024. View

20.

Trushina E, Dutta T, Persson X, Mielke M, Petersen R . Identification of altered metabolic pathways in plasma and CSF in mild cognitive impairment and Alzheimer's disease using metabolomics. PLoS One. 2013; 8(5):e63644. PMC: 3658985. DOI: 10.1371/journal.pone.0063644. View