Skip to main content
SpringerLink
Log in

Anionic metabolic profiling of urine from antibiotic-treated rats by capillary electrophoresis–mass spectrometry

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

A recently developed capillary electrophoresis (CE)-negative-ionisation mass spectrometry (MS) method was used to profile anionic metabolites in a microbial-host co-metabolism study. Urine samples from rats receiving antibiotics (penicillin G and streptomycin sulfate) for 0, 4, or 8 days were analysed. A quality control sample was measured repeatedly to monitor the performance of the applied CE-MS method. After peak alignment, relative standard deviations (RSDs) for migration time of five representative compounds were below 0.4 %, whereas RSDs for peak area were 7.9–13.5 %. Using univariate and principal component analysis of obtained urinary metabolic profiles, groups of rats receiving different antibiotic treatment could be distinguished based on 17 discriminatory compounds, of which 15 were downregulated and 2 were upregulated upon treatment. Eleven compounds remained down- or upregulated after discontinuation of the antibiotics administration, whereas a recovery effect was observed for others. Based on accurate mass, nine compounds were putatively identified; these included the microbial-mammalian co-metabolites hippuric acid and indoxyl sulfate. Some discriminatory compounds were also observed by other analytical techniques, but CE-MS uniquely revealed ten metabolites modulated by antibiotic exposure, including aconitic acid and an oxocholic acid. This clearly demonstrates the added value of CE-MS for nontargeted profiling of small anionic metabolites in biological samples.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

1H NMR:

Proton nuclear magnetic resonance

ABx-Dy:

Samples taken at day y after the use of antibiotics for x days

BGE:

Background electrolyte

CE:

Capillary electrophoresis

EOF:

Electro-osmotic flow

ESI:

Electrospray ionisation

GC:

Gas chromatography

LC:

Liquid chromatography

MS:

Mass spectrometry

NaOH:

Sodium hydroxide

PC:

Principal component

PCA:

Principal component analysis

QC:

Quality control

RPW:

Reference peak warping

RSD:

Relative standard deviation

TCA:

Tricarboxylic acid

TEA:

Triethylamine

TOF:

Time-of-flight

References

  1. Nicholson JK, Holmes E, Wilson ID (2005) Nat Rev Microbiol 3:431–438

    Article  CAS  Google Scholar 

  2. Willing BP, Russell SL, Finlay BB (2011) Nat Rev Microbiol 9:233–243

    Article  CAS  Google Scholar 

  3. van der Greef J, Stroobant P, van der Heijden R (2004) Curr Opin Chem Biol 8:559–565

    Article  Google Scholar 

  4. Fiehn O (2002) Plant Mol Biol 48:155–171

    Article  CAS  Google Scholar 

  5. Nicholson JK, Wilson ID (2003) Nat Rev Drug Discov 2:668–676

    Article  CAS  Google Scholar 

  6. Swann JR, Tuohy KM, Lindfors P, Brown DT, Wilson ID, Sidaway J, Nicholson JK, Holmes E (2011) J Proteome Res 10:3590–3603

    Article  CAS  Google Scholar 

  7. Zheng X, Xie G, Zhao A, Zhao L, Yao C, Chiu NHL, Zhou Z, Bao Y, Jia W, Nicholson JK, Jia W (2011) J Proteome Res 10:5512–5522

    Article  CAS  Google Scholar 

  8. Romick-Rosendale LE, Goodpaster AM, Hanwright PJ, Patel NB, Wheeler ET, Chona DL, Kennedy MA (2009) Magn Reson Chem 47:S36–S46

    Article  CAS  Google Scholar 

  9. Yap IKS, Li JV, Saric J, Martin F, Davies H, Wang Y, Wilson ID, Nicholson JK, Utzinger J, Marchesi JR, Holmes E (2008) J Proteome Res 7:3718–3728

    Article  CAS  Google Scholar 

  10. Williams RE, Eyton-Jones HW, Farnworth MJ, Gallagher R, Provan WM (2002) Xenobiotica 32:783–794

    Article  CAS  Google Scholar 

  11. Antunes LCM, Han J, Ferreira RBR, Lolić P, Borchers CH, Finlay BB (2011) Antimicrob Agents Chemother 55:1494–1503

    Article  CAS  Google Scholar 

  12. Ramautar R, Somsen GW, de Jong GJ (2009) Electrophoresis 30:276–291

    Article  CAS  Google Scholar 

  13. Ramautar R, Mayboroda OA, Somsen GW, de Jong GJ (2011) Electrophoresis 32:52–65

    Article  CAS  Google Scholar 

  14. Kok MGM, de Jong GJ, Somsen GW (2011) Electrophoresis 32:3016–3024

    Article  CAS  Google Scholar 

  15. Wishart DS, Knox C, Guo AC, Eisner R, Young N, Gautam B, Hau DD, Psychogios N, Dong E, Bouatra S, Mandal R, Sinelnikov I, Xia J, Jia L, Cruz JA, Lim E, Sobsey CA, Shrivastava S, Huang P, Liu P, Fang L, Peng J, Fradette R, Cheng D, Tzur D, Clements M, Lewis A, De Souza A, Zuniga A, Dawe M, Xiong Y, Clive D, Greiner R, Nazyrova A, Shaykhutdinov R, Li L, Vogel HJ, Forsythei I (2009) Nucleic Acids Res 37:D603–D610

    Article  CAS  Google Scholar 

  16. Smith CA, O’Maille G, Want EJ, Qin C, Trauger SA, Brandon TR, Custodio DE, Abagyan R, Siuzdak G (2005) Ther Drug Monit 27:747–751

    Article  CAS  Google Scholar 

  17. Horai H, Arita M, Kanaya S, Nihei Y, Ikeda T, Suwa K, Ojima Y, Tanaka K, Tanaka S, Aoshima K, Oda Y, Kakazu Y, Kusano M, Tohge T, Matsuda F, Sawada Y, Hirai MY, Nakanishi H, Ikeda K, Akimoto N, Maoka T, Takahashi H, Ara T, Sakurai N, Suzuki H, Shibata D, Neumann S, Iida T, Tanaka K, Funatsu K, Matsuura F, Soga T, Taguchi R, Saito K, Nishioka T (2010) J Mass Spectrom 45:703–714

    Article  CAS  Google Scholar 

  18. Nevedomskaya E, Derks R, Deelder AM, Mayboroda OA, Palmblad M (2009) Anal Bioanal Chem 395:2527–2533

    Article  CAS  Google Scholar 

  19. Garcia-Perez I, Vallejo M, Garcia A, Legido-Quigley C, Barbas C (2008) J Chromatogr A 1204:130–139

    Article  CAS  Google Scholar 

  20. Ramautar R, Sastre Toraño J, Somsen GW, de Jong GJ (2010) Electrophoresis 31:2319–2327

    Article  CAS  Google Scholar 

  21. Warrack BM, Hnatyshyn S, Ott K, Reily MD, Sanders M, Zhang H, Drexler DM (2009) J Chromatogr B 877:547–552

    Article  CAS  Google Scholar 

  22. Ryan D, Robards K, Prenzler PD, Kendall M (2011) Anal Chim Acta 684:17–29

    Article  CAS  Google Scholar 

  23. Swann JR, Want EJ, Geier FM, Spagou K, Wilson ID, Sidaway JE, Nicholson JK, Holmes E (2011) Proc Natl Acad Sci U S A 108:4523–4530

    Article  CAS  Google Scholar 

  24. Martin FJ, Dumas M, Wang Y, Legido-Quigley C, Yap IKS, Tang H, Zirah S, Murphy GM, Cloarec O, Lindon JC, Sprenger N, Fay LB, Kochhar S, van Bladeren P, Holmes E, Nicholson JK (2007) Mol Syst Biol 3:112

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biomolecular Analysis, Department of Pharmaceutical Sciences, Utrecht University, P.O. Box 80082, 3508 TB, Utrecht, The Netherlands

    Miranda G. M. Kok, Govert W. Somsen & Gerhardus J. de Jong

  2. Research group Analysis Techniques in Life Science, Avans Hogeschool, P.O. Box 90116, 4800 RA, Breda, The Netherlands

    Miranda G. M. Kok, Govert W. Somsen & Gerhardus J. de Jong

  3. MsMetrix, Doornhoecklaan 81, 3601 JT, Maarssen, The Netherlands

    Marco M. A. Ruijken

  4. Department of Food and Nutritional Sciences, School of Chemistry, Food and Pharmacy, University of Reading, Whiteknights, Reading, RG6 6AP, UK

    Jonathan R. Swann

  5. Department of Drug Metabolism and Pharmacokinetics IM, AstraZeneca, Alderley Park, Macclesfield, Cheshire, SK10 4TG, UK

    Ian D. Wilson

Authors
  1. Miranda G. M. Kok
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marco M. A. Ruijken
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jonathan R. Swann
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ian D. Wilson
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Govert W. Somsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gerhardus J. de Jong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Miranda G. M. Kok.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kok, M.G.M., Ruijken, M.M.A., Swann, J.R. et al. Anionic metabolic profiling of urine from antibiotic-treated rats by capillary electrophoresis–mass spectrometry. Anal Bioanal Chem 405, 2585–2594 (2013). https://doi.org/10.1007/s00216-012-6701-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-012-6701-4

Keywords

Search

Navigation