Capillary electrophoresis (CE) is well suited for the profiling of highly polar and charged metabolites as compounds are separated on the basis of their charge-to-size ratio. The protocol presented here is based on using a recently developed sheathless interfacing design, i.e., a porous tip interface, for coupling CE to electrospray ionization mass spectrometry (MS). It is demonstrated that sheathless CE-MS employing a bare fused-silica capillary at low-pH separation conditions can be used for the profiling of both cationic and anionic metabolites by only switching the MS detection and electrophoretic separation voltage polarity. The proposed sheathless CE-MS protocol allows efficient and sensitive profiles to be obtained for a broad array of charged metabolites, including amino acids, organic acids, nucleotides, and sugar phosphates, in various biological samples, such as urine and extracts of the glioblastoma cell line.
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Dr. Rawi Ramautar would like to acknowledge the financial support of the Veni and Vidi grant scheme of the Netherlands Organization for Scientific Research (NWO Veni 722.013.008 and Vidi 723.016.003).
Ramautar R, Berger R, van der Greef J et al (2013) Human metabolomics: strategies to understand biology. Curr Opin Chem Biol 17(5):841–846CrossRefGoogle Scholar
Kuehnbaum NL, Britz-McKibbin P (2013) New advances in separation science for metabolomics: resolving chemical diversity in a post-genomic era. Chem Rev 113(4):2437–2468CrossRefGoogle Scholar
Theodoridis GA, Gika HG, Want EJ et al (2012) Liquid chromatography-mass spectrometry based global metabolite profiling: a review. Anal Chim Acta 711:7–16CrossRefGoogle Scholar
Ramautar R (2016) CE-MS in metabolomics: status quo and the way forward. Bioanalysis 8(5):371–374CrossRefGoogle Scholar
Pejchinovski M, Hrnjez D, Ramirez-Torres A et al (2015) Capillary zone electrophoresis on-line coupled to mass spectrometry: a perspective application for clinical proteomics. Proteomics Clin Appl 9(5–6):453–468CrossRefGoogle Scholar
Pontillo C, Filip S, Borras DM et al (2015) CE-MS-based proteomics in biomarker discovery and clinical application. Proteomics Clin Appl 9(3–4):322–334CrossRefGoogle Scholar
Soga T, Ueno Y, Naraoka H et al (2002) Simultaneous determination of anionic intermediates for Bacillus subtilis metabolic pathways by capillary electrophoresis electrospray ionization mass spectrometry. Anal Chem 74(10):2233–2239CrossRefGoogle Scholar
Soga T, Ohashi Y, Ueno Y et al (2003) Quantitative metabolome analysis using capillary electrophoresis mass spectrometry. J Proteome Res 2(5):488–494CrossRefGoogle Scholar
Maxwell EJ, Chen DD (2008) Twenty years of interface development for capillary electrophoresis-electrospray ionization-mass spectrometry. Anal Chim Acta 627(1):25–33CrossRefGoogle Scholar
Bonvin G, Schappler J, Rudaz S (2012) Capillary electrophoresis-electrospray ionization-mass spectrometry interfaces: fundamental concepts and technical developments. J Chromatogr A 1267:17–31CrossRefGoogle Scholar
Bonvin G, Veuthey JL, Rudaz S et al (2012) Evaluation of a sheathless nanospray interface based on a porous tip sprayer for CE-ESI-MS coupling. Electrophoresis 33(4):552–562CrossRefGoogle Scholar
Ramautar R, Busnel JM, Deelder AM et al (2012) Enhancing the coverage of the urinary metabolome by sheathless capillary electrophoresis-mass spectrometry. Anal Chem 84(2):885–892CrossRefGoogle Scholar
Gulersonmez MC, Lock S, Hankemeier T et al (2016) Sheathless capillary electrophoresis-mass spectrometry for anionic metabolic profiling. Electrophoresis 37(7–8):1007–1014CrossRefGoogle Scholar
Hirayama A, Tomita M, Soga T (2012) Sheathless capillary electrophoresis-mass spectrometry with a high-sensitivity porous sprayer for cationic metabolome analysis. Analyst 137(21):5026–5033CrossRefGoogle Scholar
Moini M (2007) Simplifying CE-MS operation. 2. Interfacing low-flow separation techniques to mass spectrometry using a porous tip. Anal Chem 79(11):4241–4246CrossRefGoogle Scholar
Zhang W, Gulersonmez MC, Hankemeier T et al (2016) Sheathless capillary electrophoresis-mass spectrometry for metabolic profiling of biological samples. J Vis Exp (116). doi:https://doi.org/10.3791/54535