Abstract
Saliva is a clear, watery biofluid produced by the salivary glands to protect and lubricate the oral cavity. While mostly composed of water (99 %), the chemical composition of saliva is known to change quite dramatically in response to a variety of different physiological states, stimuli, insults and stressors. Unfortunately, among the human biofluids typically used in medical testing (such as blood and urine), saliva is rarely used. Given that saliva is the most easily accessible and readily obtained biofluid, this is rather unfortunate. Part of the reluctance to use saliva in medical testing likely has to do with the fact that its chemical composition is not well known. Here, a comprehensive characterization of the human saliva metabolome is presented. Multiple analytical platforms including nuclear magnetic resonance spectroscopy, gas chromatography mass spectrometry, direct flow injection/liquid chromatography mass spectrometry, inductively coupled plasma mass spectrometry, and high performance liquid chromatography were employed to quantify the metabolites that can be commonly detected in human saliva. Using this multiplatform approach, we were able to quantify and/or identify 308 salivary metabolites or metabolite species in human saliva. This experimental work was complemented with computer-aided literature mining that led to the identification and annotation of another 708 salivary metabolites. The combined collection of 853 non-redundant salivary metabolites or metabolite species together with their concentrations, related literature references, and links to their known disease associations are freely available at http://www.hmdb.ca/.
Similar content being viewed by others
References
Álvarez-Sánchez, B., Priego-Capote, F., & Luque de Castro, M. D. (2012). Study of sample preparation for metabolomic profiling of human saliva by liquid chromatography-time of flight/mass spectrometry. Journal of Chromatography A, 1248, 178–181.
Arakeri, G., Patil, S. G., Ramesh, D. N., Hunasgi, S., & Brennan, P. A. (2013). Evaluation of the possible role of copper ions in drinking water in the pathogenesis of oral submucous fibrosis: A pilot study. British Journal of Oral and Maxillofacial Surgery,. doi:10.1016/j.bjoms.2013.01.010.
Barbosa, F, Jr, Corrêa Rodrigues, M., Buzalaf, M., Krug, F., Gerlach, R., & Tanus-Santos, J. (2006). Evaluation of the use of salivary lead levels as a surrogate of blood lead or plasma lead levels in lead exposed subjects. Archives of Toxicology, 80(10), 633–637. doi:10.1007/s00204-006-0096-y.
Bouatra, S., Aziat, F., Mandal, R., et al. (2013). The human urine metabolome. PLoS ONE, 8(9), e73076. doi:10.1371/journal.pone.0073076.
Burt, B. A. (2006). The use of sorbitol- and xylitol-sweetened chewing gum in caries control. Journal of the American Dental Association, 137(2), 190–196.
Cámpora, P., Bermejo, A. M., Tabernero, M. J., & Fernández, P. (2003). Quantitation of cocaine and its major metabolites in human saliva using gas chromatography-positive chemical ionization-mass spectrometry (GC-PCI-MS). Journal of Analytical Toxicology, 27(5), 270–274.
Capote, F. P., Jimenez, J. R., Granados, J. M. M., & de Castro, M. D. L. (2007). Identificaion and determination of fat-soluble vitamins and metabolites in human serum by liquid chromatoghraphy/triple quadrupole mass spectrometry with multiple reaction monitoring. Rapid Communications in Mass Spectrometry, 21, 1745–1754.
Cerutti, P. A., & Trump, B. F. (1991). Inflammation and oxidative stress in carcinogenesis. Cancer Cells, 3, 1–7.
Chatzimichalakis, P. F., Samanidou, V. F., Verpoorte, R., & Papadoyannis, I. N. (2004). Development of a validated HPLC method for the determination of B-complex vitamins in pharmaceuticals and biological fluids after solid phase extraction. Journal of Separation Science, 27, 1181–1188.
Chiappin, S., Antonelli, G., Gatti, R., & De Palo, E. F. (2007). Saliva specimen: a new laboratory tool for diagnostic and basic investigation. Clinica Chimica Acta, 383(1–2), 30–40. doi:10.1016/j.cca.2007.04.011.
Cooke, M., Leeves, N., & White, C. (2003). Time profile of putrescine, cadaverine, indole and skatole in human saliva. Archives of Oral Biology, 48(4), 323–327.
Cross, S. E., Kreth, J., Wali, R. P., Sullivan, R., Shi, W., & Gimzewski, J. K. (2009). Evaluation of bacteria-induced enamel demineralization using optical profilometry. Dental Materials, 25(12), 1517–1526. doi:10.1016/j.dental.2009.07.012.
Dallmann, R., Viola, A. U., Tarokh, L., Cajochen, C., & Brown, S. A. (2012). The human circadian metabolome. Proceedings of the National Academy of Sciences, 109(7), 2625–2629. doi:10.1073/pnas.1114410109.
de Almeida Pdel, V., Gregio, A. M., Machado, M. A., de Lima, A. A., & Azevedo, L. R. (2008). Saliva composition and functions: A comprehensive review. The Journal of Contemporary Dental Practice, 9(3), 72–80.
Denny, P., Hagen, F. K, Hardt, M., et al. (2008). The proteomes of human parotid and submandibular/sublingual gland salivas collected as the ductal secretions. Journal of Proteome Research, 7(5), 1994–2006.
Distler, W., & Kroncke, A. (1981). The lactate metabolism of the oral bacterium Veillonella from human saliva. Archives of Oral Biology, 26(8), 657–661.
Fidalgo, T. K. S., Freitas-Fernandes, L. B., Angeli, R., et al. (2013). Salivary metabolite signatures of children with and without dental caries lesions. Metabolomics, 9(3), 657–666.
Fiehn, O., Wohlgemuth, G., & Scholz, M. (2005). Setup and annotation of metabolomic experiments by integrating biological and mass spectrometric metadata. In B. Ludäscher, & L. Raschid (Eds.), Data integration in the life sciences (Vol. 3615, pp. 224–239). Lecture notes in computer science. Berlin: Springer.
Fischer, D., & Ship, J. A. (1999). Effect of age on variability of parotid salivary gland flow rates over time. Age and Ageing, 28(6), 557–561.
Fiskerstrand, T., Refsum, H., Kvalheim, G., & Ueland, P. M. (1993). Homocysteine and other thiols in plasma and urine: Automated determination and sample stability. Clinical Chemistry, 39(2), 263–271.
Goldberg, S., Kozlovsky, A., Gordon, D., Gelernter, I., Sintov, A., & Rosenberg, M. (1994). Cadaverine as a putative component of oral malodor. Journal of Dental Research, 73(6), 1168–1172.
Guinan, T., Ronci, M., Kobus, H., & Voelcker, N. H. (2012). Rapid detection of illicit drugs in neat saliva using desorption/ionization on porous silicon. Talanta, 99, 791–798. doi:10.1016/j.talanta.2012.07.029.
Gwinner, W., & Gröne, H. J. (2000). Role of reactive oxygen species in glomerulonephritis. Nephrology, Dialysis, Transplantation, 15(8), 1127–1132.
Haug, K., Salek, R. M., Conesa, P., Hastings, J., de Matos, P., Rijnbeek, M., et al. (2013). MetaboLights–an open-access general-purpose repository for metabolomics studies and associated meta-data. Nucleic Acids Research, 41(Database issue), D781–D786. doi:10.1093/nar/gks1004.
Heitland, P., & Köster, H. D. (2006). Biomonitoring of 30 trace elements in urine of children and adults by ICP-MS. Clinical Chimica Acta, 365(1–2), 310–318.
Hu, G., & Sandham, H. J. (1972). Streptococcal utilization of lactic acid and its effect on pH. Archives of Oral Biology, 17(4), 729–743.
Jia, J., Sun, Y., Yang, H., et al. (2012). Effect of human saliva on wound healing. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi, 26(5), 563–566.
Kamodyova, N., Tothova, L., & Celec, P. (2013). Salivary markers of oxidative stress and antioxidant status: influence of external factors. Disease Markers, 34(5), 313–321. doi:10.3233/dma-130975.
Kaufman, E., & Lamster, I. B. (2002). The diagnostic applications of saliva—a review. Critical Reviews in Oral Biology & Medicine, 13(2), 197–212.
Kim, Y. J., Kim, Y. K., & Kho, H. S. (2010). Effects of smoking on trace metal levels in saliva. Oral Diseases, 16(8), 823–830. doi:10.1111/j.1601-0825.2010.01698.x.
Kochanska, B., Smolenski, R. T., & Knap, N. (2000). Determination of adenine nucleotides and their metabolites in human saliva. Acta Biochimica Polonica, 47(3), 877–879.
Korithoski, B., Krastel, K., & Cvitkovitch, D. G. (2005). Transport and metabolism of citrate by Streptococcus mutans. Journal of Bacteriology, 187(13), 4451–4456. doi:10.1128/jb.187.13.4451-4456.2005.
Kusmierek, K., & Bald, E. (2008). Measurement of reduced and total mercaptamine in urine using liquid chromatography with ultraviolet detection. Biomedical Chromatography, 22(4), 441–445. doi:10.1002/bmc.959.
Larsen, M. J., Jensen, A. F., Madsen, D. M., & Pearce, E. I. (1999). Individual variations of pH, buffer capacity, and concentrations of calcium and phosphate in unstimulated whole saliva. Archives of Oral Biology, 44(2), 111–117.
Lee, S., Pagoria, D., Raigrodski, A., et al. (2007). Effects of combinations of ROS scavengers on oxidative DNA damage caused by visible-light-activated camphorquinone/N, N-dimethyl-p-toluidine. Journal of Biomedical Materials Research. Part B, Applied Biomaterials, 83(2), 391–399.
Linke, H. A., Moss, S. J., Arav, L., & Chiu, P. M. (1997). Intra-oral lactic acid production during clearance of different foods containing various carbohydrates. Zeitschrift fur Ernahrungswissenschaft, 36(2), 191–197.
Magalhaes, A. C., Wiegand, A., Rios, D., Buzalaf, M. A., & Lussi, A. (2011). Fluoride in dental erosion. Monographs in Oral Science, 22, 158–170. doi:10.1159/000325167.
Mandal, R., Guo, A. C., Chaudhary, K. K., et al. (2012). Multi-platform characterization of the human cerebrospinal fluid metabolome: A comprehensive and quantitative update. Genome Medicine, 4(4), 38. doi:10.1186/gm337.
Marcotte, H., & Lavoie, M. C. (1998). Oral microbial ecology and the role of salivary immunoglobulin A. Microbiology and Molecular Biology Reviews, 62(1), 71–109.
Martin, H. J., Riazanskaia, S., & Thomas, C. L. (2012). Sampling and characterisation of volatile organic compound profiles in human saliva using a polydimethylsiloxane coupon placed within the oral cavity. Analyst, 137(16), 3627–3634. doi:10.1039/c2an35432b.
Morenkova, S. A. (2004). Comparative analysis of dependence of saliva sorbitol and fructosamine levels on blood glucose level in patients with diabetes. Biomed Khim, 50(6), 612–614.
Morris-Wiman, J., Sego, R., Brinkley, L., & Dolce, C. (2000). The effects of sialoadenectomy and exogenous EGF on taste bud morphology and maintenance. Chemical Senses, 25(1), 9–19. doi:10.1093/chemse/25.1.9.
Nakamura, Y., Kodama, H., Satoh, T., et al. (2010). Diurnal changes in salivary amino acid concentrations. Vivo, 24(6), 837–842.
Oudhoff, M. J., Bolscher, J. G. M., Nazmi, K., et al. (2008). Histatins are the major wound-closure stimulating factors in human saliva as identified in a cell culture assay. The FASEB Journal, 22(11), 3805–3812. doi:10.1096/fj.08-112003.
Park, Y. D., Jang, J. H., Oh, Y. J., & Kwon, H. J. (2014). Analyses of organic acids and inorganic anions and their relationship in human saliva before and after glucose intake. Archives of Oral Biology, 59(1), 1–11. doi:10.1016/j.archoralbio.2013.10.006.
Persson, S., Edlund, M. B., Claesson, R., & Carlsson, J. (1990). The formation of hydrogen sulfide and methyl mercaptan by oral bacteria. Oral Microbiology and Immunology, 5(4), 195–201.
Piermarini, S., Volpe, G., Federico, R., Moscone, D., & Palleschi, G. (2010). Detection of biogenic amines in human saliva using a screen-printed biosensor. Analytical Letters, 43(7–8), 1310–1316. doi:10.1080/00032710903518724.
Pittendrigh, C. S. (1993). Temporal organization: reflections of a Darwinian clock-watcher. Annual Review of Physiology, 55, 16–54. doi:10.1146/annurev.ph.55.030193.000313.
Pobozy, E., Czarkowska, W., & Trojanowicz, M. (2006). Determination of amino acids in saliva using capillary electrophoresis with fluorimetric detection. Journal of Biochemical and Biophysical Methods, 67(1), 37–47.
Psychogios, N., Hau, D. D., Peng, J., et al. (2011). The human serum metabolome. PLoS ONE, 6(2), e16957. doi:10.1371/journal.pone.0016957.
Rolla, G., Ciardi, J. E., & Bowen, W. H. (1983). Identification of IgA, IgG, lysozyme, albumin, alpha-amylase and glucosyltransferase in the protein layer adsorbed to hydroxyapatite from whole saliva. Scandinavian Journal of Dental Research, 91(3), 186–190.
Sanchez-Pablo, M. A., Gonzalez-Garcia, V., & del Castillo-Rueda, A. (2012). Study of total stimulated saliva flow and hyperpigmentation in the oral mucosa of patients diagnosed with hereditary hemochromatosis. Series of 25 cases. Medicina Oral, Patología Oral y Cirugía Bucal, 17(1), e45–e49.
Shetty, S. R., Babu, S., Kumari, S., Shetty, P., Vijay, R., & Karikal, A. (2012). Evaluation of micronutrient status in serum and saliva of oral submucous fibrosis patients: A clinicopathological study. Indian Journal of Medical and Paediatric Oncology, 33(4), 224–226. doi:10.4103/0971-5851.107087.
Silwood, C. J., Lynch, E., Claxson, A. W., & Grootveld, M. C. (2002). 1H and (13)C NMR spectroscopic analysis of human saliva. Journal of Dental Research, 81(6), 422–427.
Soini, H. A., Klouckova, I., Wiesler, D., et al. (2010). Analysis of volatile organic compounds in human saliva by a static sorptive extraction method and gas chromatography-mass spectrometry. Journal of Chemical Ecology, 36(9), 1035–1042. doi:10.1007/s10886-010-9846-7.
Spielmann, N., & Wong, D. T. (2011). Saliva: diagnostics and therapeutic perspectives. Oral Diseases, 17(4), 345–354.
Spinner, D. S., Cho, I. S., Park, S. Y., et al. (2008). Accelerated prion disease pathogenesis in Toll-like receptor 4 signaling-mutant mice. Journal of Virology, 82(21), 10701–10708. doi:10.1128/JVI.00522-08.
Sugimoto, M., Saruta, J., Matsuki, C., et al. (2013). Physiological and environmental parameters associated with mass spectrometry-based salivary metabolomic profiles. Metabolomics, 9(2), 454–463. doi:10.1007/s11306-012-0464-y.
Sugimoto, M., Wong, D. T., Hirayama, A., Soga, T., & Tomita, M. (2010). Capillary electrophoresis mass spectrometry-based saliva metabolomics identified oral, breast and pancreatic cancer-specific profiles. Metabolomics, 6(1), 78–95. doi:10.1007/s11306-009-0178-y.
Takeda, I., Stretch, C., Barnaby, P., et al. (2009). Understanding the human salivary metabolome. NMR in Biomedicine, 22(6), 577–584. doi:10.1002/nbm.1369.
Toone, R. J., Peacock, O. J., Smith, A. A., et al. (2013). Measurement of steroid hormones in saliva: Effects of sample storage condition. Scandinavian Journal of Clinical and Laboratory Investigation, 73(8), 615–621. doi:10.3109/00365513.2013.835862.
Vakkuri, O. (1985). Diurnal rhythm of melatonin in human saliva. Acta Physiologica Scandinavica, 124(3), 409–412. doi:10.1111/j.1748-1716.1985.tb07676.x.
Walsh, M. C., Brennan, L., Malthouse, J. P. G., Roche, H. M., & Gibney, M. J. (2006). Effect of acute dietary standardization on the urinary, plasma, and salivary metabolomic profiles of healthy humans. The American Journal of Clinical Nutrition, 84(3), 531–539.
Wang, D., Fan, L., Zhang, L., et al. (2012). Comparison of the total arsenic concentration between saliva and blood after oral administration of sodium arsenite to rats. Wei Sheng Yan Jiu, 41(6), 947–950.
Ward, M. E., Politzer, I. R., Laseter, J. L., & Alam, S. Q. (1976). Gas chromatographic mass spectrometric evaluation of free organic acids in human saliva. Biomedical Mass Spectrometry, 3(2), 77–80.
Wei, J., Xie, G., Zhou, Z., Shi, P., et al. (2011). Salivary metabolite signatures of oral cancer and leukoplakia. International Journal of Cancer, 129(9), 2207–2217.
Wishart, D. S., Jewison, T., Guo, A. C., et al. (2013). HMDB 3.0–The human metabolome database in 2013. Nucleic Acids Research, 41(Database issue), D801–D807. doi:10.1093/nar/gks1065.
Wishart, D. S., Lewis, M. J., Morrissey, J. A., et al. (2008). The human cerebrospinal fluid metabolome. Journal of Chromatography B, 871(2), 164–173. doi:10.1016/j.jchromb.2008.05.001.
Wisner, A., Dufour, E., Messaoudi, M., et al. (2006). Human Opiorphin, a natural antinociceptive modulator of opioid-dependent pathways. Proceedings of the National Academy of Sciences, 103(47), 17979–17984. doi:10.1073/pnas.0605865103.
Wong, D. T. (2006). Salivary diagnostics powered by nanotechnologies, proteomics and genomics. Journal of the American Dental Association, 137(3), 313–321.
Xia, Y., Peng, C., Zhou, Z., et al. (2012). Clinical significance of saliva urea, creatinine, and uric acid levels in patients with chronic kidney disease. Zhong Nan Da Xue Xue Bao Yi Xue Ban, 37(11), 1171–1176. doi:10.3969/j.issn.1672-7347.2012.11.016.
Zappacosta, B., Manni, A., Persichilli, S., et al. (2003). HPLC analysis of some sulphur compounds in saliva: Comparison between healthy subjects and periodontopathic patients. Clinica Chimica Acta, 338(1–2), 57–60.
Zheng, J., Dixon, R. A., & Li, L. (2012). Development of isotope labeling LC-MS for human salivary metabolomics and application to profiling metabolome changes associated with mild cognitive impairment. Analytical Chemistry, 84(24), 10802–10811. doi:10.1021/ac3028307.
Funding
Funding for this research has been provided by Genome Canada, Genome Alberta, The Canadian Institutes of Health Research, Alberta Innovates, The National Research Council and The National Institute of Nanotechnology. The funders had no role in study design, data collection. data analysis, decision to publish, or preparation of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
All authors declare that they have no conflict of interest.
Ethical approval
The study complied with all applicable institutional guidelines and terms of the Declaration of Helsinki of 1975 (as revised in 2008) for investigation of human subjects. The research involving human subjects was based on their informed consent. All participants agreed to participate in this study and to contribute saliva samples for metabolomic analysis. All samples were collected in accordance with the ethical guidelines mandated by the University of Alberta as approved by the University’s Health Research Ethics Board.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Dame, Z.T., Aziat, F., Mandal, R. et al. The human saliva metabolome. Metabolomics 11, 1864–1883 (2015). https://doi.org/10.1007/s11306-015-0840-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11306-015-0840-5