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. 2013 Sep 4;8(9):e73076.
doi: 10.1371/journal.pone.0073076. eCollection 2013.

The Human Urine Metabolome

Free PMC article

The Human Urine Metabolome

Souhaila Bouatra et al. PLoS One. .
Free PMC article


Urine has long been a "favored" biofluid among metabolomics researchers. It is sterile, easy-to-obtain in large volumes, largely free from interfering proteins or lipids and chemically complex. However, this chemical complexity has also made urine a particularly difficult substrate to fully understand. As a biological waste material, urine typically contains metabolic breakdown products from a wide range of foods, drinks, drugs, environmental contaminants, endogenous waste metabolites and bacterial by-products. Many of these compounds are poorly characterized and poorly understood. In an effort to improve our understanding of this biofluid we have undertaken a comprehensive, quantitative, metabolome-wide characterization of human urine. This involved both computer-aided literature mining and comprehensive, quantitative experimental assessment/validation. The experimental portion employed NMR spectroscopy, gas chromatography mass spectrometry (GC-MS), direct flow injection mass spectrometry (DFI/LC-MS/MS), inductively coupled plasma mass spectrometry (ICP-MS) and high performance liquid chromatography (HPLC) experiments performed on multiple human urine samples. This multi-platform metabolomic analysis allowed us to identify 445 and quantify 378 unique urine metabolites or metabolite species. The different analytical platforms were able to identify (quantify) a total of: 209 (209) by NMR, 179 (85) by GC-MS, 127 (127) by DFI/LC-MS/MS, 40 (40) by ICP-MS and 10 (10) by HPLC. Our use of multiple metabolomics platforms and technologies allowed us to identify several previously unknown urine metabolites and to substantially enhance the level of metabolome coverage. It also allowed us to critically assess the relative strengths and weaknesses of different platforms or technologies. The literature review led to the identification and annotation of another 2206 urinary compounds and was used to help guide the subsequent experimental studies. An online database containing the complete set of 2651 confirmed human urine metabolite species, their structures (3079 in total), concentrations, related literature references and links to their known disease associations are freely available at

Conflict of interest statement

Competing Interests: Ralf Bogumil and Cornelia Roehring are employed by BIOCRATES Life Sciences AG. This company produces kits for targeted metabolomic analyses. This does not alter the authors’ adherence to all the PLOS ONE policies on sharing data and materials.


Figure 1
Figure 1. Typical 500 MHz 1H-NMR spectra of urine from human urine.
Numbers indicates the following metabolites: 1: creatinine; 2: citric acid; 3: glycine; 4: formic acid; 5: methanol; 6: guanidoacetic acid; 7: acetic acid; 8: L-cysteine; 9: glycolic acid; 10: creatine; 11: isocitric acid; 12: hippuric acid; 13: L-glutamine; 14: L-alanine; 15: L-lysine; 16: gluconic acid; 17∶2- hydroxyglutaric acid; 18: D-glucose; 19: indoxyl sulfate; 20: trimethyl-N-oxide; 21: ethanolamine; 22: L-lactic acid; 23: taurine; 24: L-threonine; 25: dimethylamine; 26: pyroglutamic acid; 27: trigonelline; 28: sucrose; 29: trimethylamine; 30: mannitol; 31: L-serine; 32: acetone; 33: L-cystine; 34: adipic acid; 35: L-histidine; 36: L-tyrosine; 37: imidazole; 38: mandelic acid; 39: dimethylglycine; 40: Cis-aconitic acid; 41: urea; 42∶3-(3-hydroxyphenyl)-3-hydroxypropanoic acid (HPHPA); 43: phenol; 45: isobutyric acid; 46: methylsuccinic acid; 47∶3-aminoisobutyric acid; 48: L-fucose; 49: N-acetylaspartic acid; 50: N-acetylneuraminic acid; 51: acetoacetic acid; 52: Alpha-aminoadipic acid; 53: methylguanidine; 54: phenylacetylglutamine.
Figure 2
Figure 2. Typical total ion chromatogram of water-soluble metabolites extracted from human urine.
Numbers indicate the following metabolites (quantified compounds): 1: L-valine; 2: oxalic acid; 3∶3-hydroxyisobutyric acid; 4: L-serine; 5: glycine; 6: succinic acid; 7: glyceric acid; 8∶4-deoxythreonic acid; 9∶2,4-dihydroxybutanoic acid; 10∶3,4-dihydroxybutanoic acid; 11: adipic acid; 12: creatinine; 13: threonic acid; 14: L-phenylalanine; 15: p-hydroxyphenylacetic acid; 16: L-ornithine; 17: L-asparagine; 18: L-arabinose; 19: D-xylitol; 20: D-xylulose; 21: Cis/Trans-aconitic acid; 22: hippuric acid; 23: isocitric acid; 24: D-galactose; 25: D-glucose; 26: L- tyrosine; 27: sorbitol; 28: gluconic acid; 29: scyllitol; 30: myoinositol; 31: uric acid; 32: pseudouridine.
Figure 3
Figure 3. Typical GC-MS total ion chromatogram of organic acids extracted from human urine.
Numbers indicate the following metabolites (quantified compounds): 1: pyruvic acid; 2: L-lactic acid; 3: alpha-hydroxyisobutyric acid; 4: glycolic acid; 5: levulinic acid; 6∶3-hydroxyisovaleric acid; 7∶2-hydroxy-2-methylbutyric acid; 8: hydroxypropionic acid; 9∶2-methyl-3-hydroxybutyric acid; 10: malonic acid; 11∶3-hydroxyisovaleric acid; 12: methylmalonic acid; 13∶2-ethylhydracrylic acid; 14: benzoic acid; 15: phosphoric acid; 16: ethylmalonic acid; 17: succinic acid; 18: methylsuccinic acid; 19∶4-deoxythreonic acid; 20∶5-hydroxyhexanoic acid; 21: citraconic acid; 22: glutaric acid; 23: m-chlorobenzoic acid; 24; 3,4-dihydroxybutanoic acid; 25∶3-methylglutaconic acid; 26: adipic acid; 27: pyroglutamic acid; 28∶3-methyladipic acid; 29: sumiki’s acid; 30: o-hydroxyphenylacetic acid; 31: oxoglutaric acid; 32: pimelic acid; 33∶3-hydroxymethylglutaric acid; 34∶3-hydroxyphenylacetic acid; 35∶4-hydroxybenzoic acid; 36∶2-furoylglycine; 37: suberic acid; 38: quinolinic acid; 39: Cis/Trans-aconitic acid; 40: homovanillic acid; 41: azelaic acid; 42: hippuric acid; 43∶3,4-dihydroxybenzeneacetic acid; 44∶3-(3-hydroxyphenyl)-3-hydroxypropanoic acid (HPHPA); 45: vanillylmandelic acid; 46∶4-hydroxyphenyllactic acid; 47: indoleacetic acid; 48: palmitic acid; 49: kynurenic acid; 50∶3-hydroxyhippuric acid; 51∶3-hydroxysebacic acid; 52: Trans-ferulic acid; 53∶5-hydroxyindoleacetic acid; 54: stearic acid.
Figure 4
Figure 4. Typical gas chromatogram of volatile organic compounds (VOC) from a pooled human urine sample.
Numbers indicate the following metabolites: 1∶3-methyl sulfolane; 2∶3-hexanone; 3∶2-pentanone; 4∶1-hydroxy-2-pentanone; 5: allyl methylsulphide; 6: dimethyl disulfide; 7∶4-heptanone; 8∶1-methylcyclohexanol; 9; 2-hexanone; 10∶3,4-dimethylthiophene; 11: diallyl sulphide; 12∶5-methyl-2-hexanone; 13∶1,3-dithio cyclohexane; 14: dimethyl trisulfide; 15: phenol; 16: o-cymene; 17: p-cymene; 18: m-cymene; 19∶1,4-cineol; 20: p-cresol; 21: linalool oxide; 22: iso-menthol; 23: Alpha-p-dimethylstyrene; 24: L-menthol; 25: undecane; 26: ledene oxide (II); 27: salicylic acid methyl ester; 28: Beta-carvone; 29: piperitone; 30: o-thymol; 31: Beta-cyclocitrol; 32∶4,7-dimethyl-benzofuran; 33: cuminal: 34∶2,6,10,10 Tetramethyl-oxa-spiro-4,5-dec-6-ene; 35∶4-(2,6,6-trimethyl-1-cyclohexen-1-yl)-2-butanone; 36∶1,2,3,4-tetrahydro-1,1,6-trimethyl naphthalene; 37: Alpha-cedrene; 38∶1,2,3,4-tetrahydro-1,5,7-trimethylnapthalene; 39∶1,2-diydro-1,1,6-trimethyl-napthalene; 40: Beta-guaiene; 41: Beta-damascenone; 42∶2,5-cyclohexadiene-1,4-dione-2,6-di-tert-butyl; 43: himachalene; 44∶4-(2,6,6-trimethylcyclohexa-1,3-dienyl)-but-3-en-2-one: 45∶1-(2,6,6-trimethyl-1-cyclohexen-1-yl)-1-penten-3-one; 46∶2,4-Bis (1,1-dimethylethyl)-phenol; 47∶1-(2,3,6-trimethyl phenyl)-3-buten-2-one; 48: L-calamenene; 49: Beta-vatirenene; 50∶1,6,7-trimethylnaphthalene; 51: azulol; 52∶3,3,5,6-tetramethyl-1-indanone.
Figure 5
Figure 5. Venn diagram showing the overlap of urine metabolites detected by NMR, GC-MS, DFI/LC-MS/MS, ICP-MS, HPLC coupled to UV detection and HPLC coupled to fluorescence detection compared to the detectable urine metabolome.

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    1. Bundy JG, Davey MP, Viant MR (2009) Environmental metabolomics: a critical review and future perspectives. Metabolomics 5: 3–21.
    1. Montoliu I, Genick U, Ledda M, Collino S, Martin FP, et al. (2013) Current status on genome-metabolome-wide associations: an opportunity in nutrition research. Genes Nutr 8: 19–27. - PMC - PubMed
    1. Nicholson JK, Holmes E, Kinross JM, Darzi AW, Takats Z, et al. (2012) Metabolic phenotyping in clinical and surgical environments. Nature 491: 384–392. - PubMed
    1. Viant MR (2008) Recent developments in environmental metabolomics. Mol Biosyst 4: 980–986. - PubMed
    1. Brindle JT, Antti H, Holmes E, Tranter G, Nicholson JK, et al. (2003) Rapid and noninvasive diagnosis of the presence and severity of coronary heart disease using 1H-NMR-based metabonomics. Nat Med 9: 477–477. - PubMed

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