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

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

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.

Animals, Anti-Bacterial Agents/metabolism, Electrophoresis, Capillary/methods, Mass Spectrometry/methods, Metabolome, Penicillin G/metabolism, Rats, Streptomycin/metabolism
1618-2642
2585-2594
Kok, Miranda G M
f0acf585-7a72-4e73-92c3-0c318f8a4bd1
Ruijken, Marco M A
efbef27b-850f-4b91-ba8f-991a1400625f
Swann, Jonathan R
7c11a66b-f4b8-4dbf-aa17-ad8b0561b85c
Wilson, Ian D
d7da811b-6cc9-4166-82d0-e780c95122d2
Somsen, Govert W
350a8603-2ed1-4096-ae44-daa9ee47d850
de Jong, Gerhardus J
526765f9-3c03-4bba-968e-e944bd2a49d1
Kok, Miranda G M
f0acf585-7a72-4e73-92c3-0c318f8a4bd1
Ruijken, Marco M A
efbef27b-850f-4b91-ba8f-991a1400625f
Swann, Jonathan R
7c11a66b-f4b8-4dbf-aa17-ad8b0561b85c
Wilson, Ian D
d7da811b-6cc9-4166-82d0-e780c95122d2
Somsen, Govert W
350a8603-2ed1-4096-ae44-daa9ee47d850
de Jong, Gerhardus J
526765f9-3c03-4bba-968e-e944bd2a49d1

Kok, Miranda G M, Ruijken, Marco M A, Swann, Jonathan R, Wilson, Ian D, Somsen, Govert W and de Jong, Gerhardus J (2013) Anionic metabolic profiling of urine from antibiotic-treated rats by capillary electrophoresis-mass spectrometry. Analytical and Bioanalytical Chemistry, 405 (8), 2585-2594. (doi:10.1007/s00216-012-6701-4).

Record type: Article

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.

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More information

e-pub ahead of print date: 12 January 2013
Published date: March 2013
Keywords: Animals, Anti-Bacterial Agents/metabolism, Electrophoresis, Capillary/methods, Mass Spectrometry/methods, Metabolome, Penicillin G/metabolism, Rats, Streptomycin/metabolism

Identifiers

Local EPrints ID: 439402
URI: http://eprints.soton.ac.uk/id/eprint/439402
ISSN: 1618-2642
PURE UUID: debaa92d-13b1-44d3-b7f1-4875194bd8f4
ORCID for Jonathan R Swann: ORCID iD orcid.org/0000-0002-6485-4529

Catalogue record

Date deposited: 21 Apr 2020 16:52
Last modified: 17 Mar 2024 04:00

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Contributors

Author: Miranda G M Kok
Author: Marco M A Ruijken
Author: Ian D Wilson
Author: Govert W Somsen
Author: Gerhardus J de Jong

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