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

Analytical and bioanalytical chemistry (2013-01-15)
Miranda G M Kok, Marco M A Ruijken, Jonathan R Swann, Ian D Wilson, Govert W Somsen, Gerhardus J de Jong
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.

MATERIALS
Product Number
Brand
Product Description

Supelco
Penicillin G potassium salt, VETRANAL®, analytical standard
Sigma-Aldrich
Penicillin G potassium salt, 95.0 - 102.0%
Sigma-Aldrich
Penicillin G potassium salt
Sigma-Aldrich
Penicillin G potassium salt, powder, BioReagent, suitable for cell culture
Supelco
Streptomycin solution, ~1 mg/mL in 1 mM EDTA, analytical standard
Sigma-Aldrich
Streptomycin sulfate salt, powder, BioXtra, suitable for mouse embryo cell culture
Sigma-Aldrich
Streptomycin sulfate salt, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Streptomycin sulfate salt, powder
Sigma-Aldrich
Penicillin G sodium salt, ~1650 U/mg
Sigma-Aldrich
Penicillin G sodium salt, powder, BioReagent, suitable for cell culture
Supelco
Streptomycin sesquisulfate hydrate, VETRANAL®, analytical standard
Sigma-Aldrich
Penicillin G sodium salt, 96.0-102.0%