Metabolic Profiling of Urine by Capillary Electrophoresis-Mass Spectrometry Using Non-covalently Coated Capillaries

  • Rawi RamautarEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1730)


In the field of metabolomics, capillary electrophoresis-mass spectrometry (CE-MS) can be considered a very useful analytical tool for the profiling of polar and charged metabolites. However, variability of migration time is an important issue in CE. An elegant way to minimize this problem is the use of non-covalently coated capillaries that is dynamic coating of the bare fused-silica capillary with solutions of charged polymers. In this protocol, an improved strategy for the profiling of cationic metabolites in urine by CE-MS using multilayered non-covalent capillary coatings is presented. Capillaries are coated with a bilayer of polybrene (PB) and poly(vinyl sulfonate) (PVS) or with a triple layer of PB, dextran sulfate (DS), and PB. The bilayer- and triple-layer-coated capillaries have a negative and positive outside layer, respectively. It is shown that the use of such capillaries provides very repeatable migration times.

Key words

Capillary electrophoresis Mass spectrometry Metabolomics Cationic metabolites Non-covalently coated capillaries Urine 



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).


  1. 1.
    Ramautar R, Somsen GW, de Jong GJ (2013) The role of CE–MS in metabolomics. In: Metabolomics in practice. Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany, pp 177–208. Scholar
  2. 2.
    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):2437–2468. Scholar
  3. 3.
    Hirayama A, Wakayama M, Soga T (2014) Metabolome analysis based on capillary electrophoresis-mass spectrometry. TrAC Trends Anal Chem 61:215–222. Scholar
  4. 4.
    Ramautar R (2016) Capillary electrophoresis-mass spectrometry for clinical metabolomics. Adv Clin Chem 74:1–34. Scholar
  5. 5.
    Nevedomskaya E, Derks R, Deelder AM, Mayboroda OA, Palmblad M (2009) Alignment of capillary electrophoresis-mass spectrometry datasets using accurate mass information. Anal Bioanal Chem 395(8):2527–2533. Scholar
  6. 6.
    Kok MG, Ruijken MM, Swann JR, Wilson ID, Somsen GW, de Jong GJ (2013) Anionic metabolic profiling of urine from antibiotic-treated rats by capillary electrophoresis-mass spectrometry. Anal Bioanal Chem 405(8):2585–2594. Scholar
  7. 7.
    Garcia-Perez I, Whitfield P, Bartlett A, Angulo S, Legido-Quigley C, Hanna-Brown M, Barbas C (2008) Metabolic fingerprinting of Schistosoma Mansoni infection in mice urine with capillary electrophoresis. Electrophoresis 29(15):3201–3206. Scholar
  8. 8.
    Huhn C, Ramautar R, Wuhrer M, Somsen GW (2010) Relevance and use of capillary coatings in capillary electrophoresis-mass spectrometry. Anal Bioanal Chem 396(1):297–314. Scholar
  9. 9.
    Dominguez-Vega E, Haselberg R, Somsen GW (2016) Capillary zone electrophoresis-mass spectrometry of intact proteins. Methods Mol Biol 1466:25–41. Scholar
  10. 10.
    Katayama H, Ishihama Y, Asakawa N (1998) Stable cationic capillary coating with successive multiple ionic polymer layers for capillary electrophoresis. Anal Chem 70(24):5272–5277CrossRefPubMedGoogle Scholar
  11. 11.
    Catai JR, Torano JS, de Jong GJ, Somsen GW (2006) Efficient and highly reproducible capillary electrophoresis-mass spectrometry of peptides using Polybrene-poly(vinyl sulfonate)-coated capillaries. Electrophoresis 27(11):2091–2099. Scholar
  12. 12.
    Soga T, Ueno Y, Naraoka H, Ohashi Y, Tomita M, Nishioka T (2002) Simultaneous determination of anionic intermediates for Bacillus Subtilis metabolic pathways by capillary electrophoresis electrospray ionization mass spectrometry. Anal Chem 74(10):2233–2239CrossRefPubMedGoogle Scholar
  13. 13.
    Ramautar R, Torano JS, Somsen GW, de Jong GJ (2010) Evaluation of CE methods for global metabolic profiling of urine. Electrophoresis 31(14):2319–2327. Scholar

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.Division of Systems Biomedicine and Pharmacology, Leiden Academic Center for Drug ResearchLeiden UniversityLeidenThe Netherlands

Personalised recommendations