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Comparison of capillary electrophoresis-mass spectrometry and hydrophilic interaction chromatography-mass spectrometry for anionic metabolic profiling of urine.
Talanta. 2015 Jan; 132:1-7.T

Abstract

In order to assess the utility of a recently developed capillary electrophoresis-mass spectrometry (CE-MS) method for the study of anionic metabolites in urine, a comparison was made with hydrophilic interaction chromatography-MS (HILIC-MS) using negative electrospray ionization. After optimization of the HILIC conditions, a gradient employing 10mM ammonium acetate (pH 6.8) in acetonitrile-water (5 min 90% acetonitrile followed by 90%-50% acetonitrile in 10 min) was selected, providing baseline separation of five representative anionic test metabolites. Relative standard deviations (RSDs) for HILIC retention times and peak areas were below 0.2% and 7.7%, respectively, and detection limits were in the range 0.04-2.21 μM. Metabolites in rat urine could also be analysed in a reproducible way with retention time and peak area RSDs below 0.6% and 13.6%, respectively. The CE-MS and HILIC-MS methods were compared in terms of reproducibility, sensitivity, selectivity and coverage of the anionic urinary metabolome. In general, peak area RSDs were similar whereas HILIC-MS yielded better retention-time repeatability and up to 80 times lower detection limits (expressed in injected concentration) for test metabolites as compared to CE-MS. Rat urine analysis by HILIC-MS provided detection of 1360 molecular features compared to 347 molecular features revealed with CE-MS. Of these, a number of 144 molecular features were found with both HILIC-MS and CE-MS, which showed on average 10 times higher peak areas in HILIC-MS. The HILIC retention and CE migration times of the common features were clearly not correlated. The HILIC and CE behavior of the test metabolites and 16 putatively identified common features were evaluated involving their physicochemical properties, indicating a markedly different separation selectivity, and thus significant degree of orthogonality of HILIC and CE.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Kok MG
Biomolecular Analysis, Department of Pharmaceutical Sciences, Utrecht University, P.O. Box 80082, 3508 TB Utrecht, The Netherlands; Research group Analysis Techniques in Life Sciences, Avans Hogeschool, P.O. Box 90116, 4800 RA Breda, The Netherlands. Electronic address: M.G.M.Kok@uu.nl.
Somsen GW
Biomolecular Analysis, Department of Pharmaceutical Sciences, Utrecht University, P.O. Box 80082, 3508 TB Utrecht, The Netherlands; AIMSS Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, VU University, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
de Jong GJ
Biomolecular Analysis, Department of Pharmaceutical Sciences, Utrecht University, P.O. Box 80082, 3508 TB Utrecht, The Netherlands; Research group Analysis Techniques in Life Sciences, Avans Hogeschool, P.O. Box 90116, 4800 RA Breda, The Netherlands.

MeSH

AcetatesAcetonitrilesAnimalsAnionsElectrophoresis, CapillaryHydrophobic and Hydrophilic InteractionsLimit of DetectionMass SpectrometryMetabolomeRatsReference StandardsSpectrometry, Mass, Electrospray IonizationUrinalysisWater

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

25476271

Citation

Kok, Miranda G M., et al. "Comparison of Capillary Electrophoresis-mass Spectrometry and Hydrophilic Interaction Chromatography-mass Spectrometry for Anionic Metabolic Profiling of Urine." Talanta, vol. 132, 2015, pp. 1-7.
Kok MG, Somsen GW, de Jong GJ. Comparison of capillary electrophoresis-mass spectrometry and hydrophilic interaction chromatography-mass spectrometry for anionic metabolic profiling of urine. Talanta. 2015;132:1-7.
Kok, M. G., Somsen, G. W., & de Jong, G. J. (2015). Comparison of capillary electrophoresis-mass spectrometry and hydrophilic interaction chromatography-mass spectrometry for anionic metabolic profiling of urine. Talanta, 132, 1-7. https://doi.org/10.1016/j.talanta.2014.08.047
Kok MG, Somsen GW, de Jong GJ. Comparison of Capillary Electrophoresis-mass Spectrometry and Hydrophilic Interaction Chromatography-mass Spectrometry for Anionic Metabolic Profiling of Urine. Talanta. 2015;132:1-7. PubMed PMID: 25476271.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparison of capillary electrophoresis-mass spectrometry and hydrophilic interaction chromatography-mass spectrometry for anionic metabolic profiling of urine. AU - Kok,Miranda G M, AU - Somsen,Govert W, AU - de Jong,Gerhardus J, Y1 - 2014/08/27/ PY - 2014/03/20/received PY - 2014/08/13/revised PY - 2014/08/18/accepted PY - 2014/12/6/entrez PY - 2014/12/6/pubmed PY - 2015/7/29/medline KW - Anionic metabolic profiling KW - Capillary electrophoresis KW - Hydrophilic interaction chromatography KW - Mass spectrometry KW - Urine SP - 1 EP - 7 JF - Talanta JO - Talanta VL - 132 N2 - In order to assess the utility of a recently developed capillary electrophoresis-mass spectrometry (CE-MS) method for the study of anionic metabolites in urine, a comparison was made with hydrophilic interaction chromatography-MS (HILIC-MS) using negative electrospray ionization. After optimization of the HILIC conditions, a gradient employing 10mM ammonium acetate (pH 6.8) in acetonitrile-water (5 min 90% acetonitrile followed by 90%-50% acetonitrile in 10 min) was selected, providing baseline separation of five representative anionic test metabolites. Relative standard deviations (RSDs) for HILIC retention times and peak areas were below 0.2% and 7.7%, respectively, and detection limits were in the range 0.04-2.21 μM. Metabolites in rat urine could also be analysed in a reproducible way with retention time and peak area RSDs below 0.6% and 13.6%, respectively. The CE-MS and HILIC-MS methods were compared in terms of reproducibility, sensitivity, selectivity and coverage of the anionic urinary metabolome. In general, peak area RSDs were similar whereas HILIC-MS yielded better retention-time repeatability and up to 80 times lower detection limits (expressed in injected concentration) for test metabolites as compared to CE-MS. Rat urine analysis by HILIC-MS provided detection of 1360 molecular features compared to 347 molecular features revealed with CE-MS. Of these, a number of 144 molecular features were found with both HILIC-MS and CE-MS, which showed on average 10 times higher peak areas in HILIC-MS. The HILIC retention and CE migration times of the common features were clearly not correlated. The HILIC and CE behavior of the test metabolites and 16 putatively identified common features were evaluated involving their physicochemical properties, indicating a markedly different separation selectivity, and thus significant degree of orthogonality of HILIC and CE. SN - 1873-3573 UR - https://www.unboundmedicine.com/medline/citation/25476271/Comparison_of_capillary_electrophoresis_mass_spectrometry_and_hydrophilic_interaction_chromatography_mass_spectrometry_for_anionic_metabolic_profiling_of_urine_ DB - PRIME DP - Unbound Medicine ER -