title: Hydrophilic interaction chromatography-mass spectrometry for anionic metabolic profiling of urine from antibiotic-treated rats.
abstract: Hydrophilic interaction chromatography-mass spectrometry (HILIC-MS) was used for anionic metabolic profiling of urine from antibiotic-treated rats to study microbial-host co-metabolism. Rats were treated with the antibiotics penicillin G and streptomycin sulfate for four or eight days and compared to a control group. Urine samples were collected at day zero, four and eight, and analyzed by HILIC-MS. Multivariate data analysis was applied to the urinary metabolic profiles to identify biochemical variation between the treatment groups. Principal component analysis found a clear distinction between those animals receiving antibiotics and the control animals, with twenty-nine discriminatory compounds of which twenty were down-regulated and nine up-regulated upon treatment. In the treatment group receiving antibiotics for four days, a recovery effect was observed for seven compounds after cessation of antibiotic administration. Thirteen discriminatory compounds could be putatively identified based on their accurate mass, including aconitic acid, benzenediol sulfate, ferulic acid sulfate, hippuric acid, indoxyl sulfate, penicillin G, phenol and vanillin 4-sulfate. The rat urine samples had previously been analyzed by capillary electrophoresis (CE) with MS detection and proton nuclear magnetic resonance ((1)H NMR) spectroscopy. Using CE-MS and (1)H NMR spectroscopy seventeen and twenty-five discriminatory compounds were found, respectively. Both hippuric acid and indoxyl sulfate were detected across all three platforms. Additionally, eight compounds were observed with both HILIC-MS and CE-MS. Overall, HILIC-MS appears to be highly complementary to CE-MS and (1)H NMR spectroscopy, identifying additional compounds that discriminate the urine samples from antibiotic-treated and control rats.
authors: Biomolecular Analysis, Department of Pharmaceutical Sciences, Utrecht University, P.O. Box 80082, 3508 TB Utrecht, The Netherlands; Research group Analysis Techniques in Life Sciences, Avans Hogeschool, P.O. Box 90116, 4800 RA Breda, The Netherlands. Electronic address: M.G.M.Kok@uu.nl.
authors: Miranda G M
authors: MG
authors: Kok
authors: Department of Food and Nutritional Sciences, School of Chemistry, Food and Pharmacy, University of Reading, Whiteknights, Reading RG6 6AP, United Kingdom.
authors: Jonathan R
authors: JR
authors: Swann
authors: Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, South Kensington, London SW7 2AZ, United Kingdom.
authors: Ian D
authors: ID
authors: Wilson
authors: AIMSS Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, VU University, de Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
authors: Govert W
authors: GW
authors: Somsen
authors: Biomolecular Analysis, Department of Pharmaceutical Sciences, Utrecht University, P.O. Box 80082, 3508 TB Utrecht, The Netherlands; Research group Analysis Techniques in Life Sciences, Avans Hogeschool, P.O. Box 90116, 4800 RA Breda, The Netherlands.
authors: Gerhardus J
authors: GJ
authors: de Jong
mesh_terms: D000818:Animals; D000838:Anions / Q000737:chemistry* / Q000652:urine*; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002853:Chromatography, Liquid / Q000379:methods; D019075:Electrophoresis, Capillary / Q000379:methods; D006626:Hippurates / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D055442:Metabolome / Q000187:drug effects; D055432:Metabolomics / Q000379:methods; D010400:Penicillin G / Q000737:chemistry; D025341:Principal Component Analysis / Q000379:methods; D051381:Rats; D017208:Rats, Wistar; D013307:Streptomycin / Q000737:chemistry; D014556:Urine / Q000737:chemistry*
chemical_list: D000838:Anions; D000900:Anti-Bacterial Agents; D006626:Hippurates; D010400:Penicillin G; C030514:hippuric acid; D013307:Streptomycin
keywords: Anionic metabolic profiling; Antibiotics; Gut microbiota; Hydrophilic interaction chromatography; Mass spectrometry