The Metabolomic Analysis of Human Milk Offers Unique Insights into Potential Child Health Benefits


Purpose of Review

Human milk is the gold standard of infant nutrition. The milk changes throughout lactation and is tailored for the infant providing the nutrients, minerals and vitamins necessary for supporting healthy infant growth. Human milk also contains low molecular weight compounds (metabolites) possibly eliciting important bioactivity. Metabolomics is the study of these metabolites. The purpose of this review was to examine recent metabolomics studies and cohort studies on human milk to assess the impact of human milk metabolomic analyses combined with investigations of infant growth and development.

Recent Findings

The metabolite profile of human milk varies among other factors according to lactation stage, gestation at birth, and maternal genes, diet and disease state. Few studies investigate how these variations impact infant growth and development.


Several time-related factors affecting human milk metabolome are potentially ubiquitous among mothers, although maternal-related factors are heavily confounded, which complicates studies of metabolite abundancies and variabilities and further possibilities of observing cause and effect in infants.

This is a preview of subscription content, access via your institution.


Papers of particular interest, published recently, have been highlighted as: • Of importance

  1. 1.

    Ballard O, Morrow AL. Human milk composition. Nutrients and Bioactive Factors. Pediatr Clin N Am. 2013;60:49–74.

    Article  Google Scholar 

  2. 2.

    Wopereis H, Oozeer R, Knipping K, Belzer C, Knol J. The first thousand days—intestinal microbiology of early life: establishing a symbiosis. Pediatr Allergy Immunol. 2014;25:428–38.

    Article  Google Scholar 

  3. 3.

    Pacheco AR, Barile D, Underwood MA, Mills DA. The impact of the milk glycobiome on the neonate gut microbiota. Annu Rev Anim Biosci. 2015;3:419–45.

    CAS  Article  Google Scholar 

  4. 4.

    Newburg DS, Ruiz-Palacios GM, Morrow AL. Human milk glycans protect infants against enteric pathogens. Annu Rev Nutr. 2005;25:37–58.

    CAS  Article  Google Scholar 

  5. 5.

    Kunz C, Rudloff S, Baier W. Oligosaccharides in human milk: structural, functional, and metabolic aspects. Annu Rev Nutr. 2000;20:699–722.

    CAS  Article  Google Scholar 

  6. 6.

    • Ten-Doménech I, Ramos-Garcia V, Piñeiro-Ramos JD, Gormaz M, Parra-Llorca A, Vento M, et al. Current practice in untargeted human milk metabolomics. Metabolites. 2020;10:43 Review on methodological considerations when investigating human milk metabolome. This review highlights distinct factors related to study protocol and design that combined influence the metabolomic outcome. Furthermore, the authors stress the need of a multi-omics approach and quality control.

    Article  Google Scholar 

  7. 7.

    Sundekilde UK, Downey E, O’Mahony J, O’Shea C-A, Ryan C, Kelly A, et al. The effect of gestational and lactational age on the human milk metabolome. Nutrients. 2016;8:304.

    Article  Google Scholar 

  8. 8.

    Spevacek AR, Smilowitz JT, Chin EL, Underwood MA, German JB, Slupsky CM. Infant maturity at birth reveals minor differences in the maternal milk metabolome in the first month of lactation. J Nutr. 2015;145:1698–708.

    CAS  Article  Google Scholar 

  9. 9.

    Shoji H, Shimizu T. Effect of human breast milk on biological metabolism in infants. Pediatr Int. 2019;61:6–15.

    CAS  Article  Google Scholar 

  10. 10.

    Martin FP, Sprenger N, Montoliu I, Rezzi S, Kochhar S, Nicholson JK. Dietary modulation of gut functional ecology studied by fecal metabonomics. J Proteome Res. 2010;9:5284–95.

    CAS  Article  Google Scholar 

  11. 11.

    Nicholson JK, Holmes E, Kinross J, Burcelin R, Gibson G, Jia W, et al. Host-gut microbiota metabolic interactions. Science (80- ). 2012;336:1262–7.

    CAS  Article  Google Scholar 

  12. 12.

    Urbaniak C, McMillan A, Angelini M, Gloor GB, Sumarah M, Burton JP, et al. Effect of chemotherapy on the microbiota and metabolome of human milk, a case report. Microbiome. 2014;2:24.

    Article  PubMed  PubMed Central  Google Scholar 

  13. 13.

    Isganaitis E, Venditti S, Matthews TJ, Lerin C, Demerath EW, Fields DA. Maternal obesity and the human milk metabolome: associations with infant body composition and postnatal weight gain. Am J Clin Nutr. 2019;110:111–20.

    Article  Google Scholar 

  14. 14.

    Andreas NJ, Hyde MJ, Gomez-Romero M, Lopez-Gonzalvez MA, Villaseñor A, Wijeyesekera A, et al. Multiplatform characterization of dynamic changes in breast milk during lactation. Electrophoresis. 2015;36:2269–85.

    CAS  Article  Google Scholar 

  15. 15.

    Wishart DS. Exploring the human metabolome by nuclear magnetic resonance spectroscopy and mass spectrometry. In: Lutz N, Sweedler J, Wevers R, editors. Methodol. Metabolomics. Cambridge: Cambridge University Press; 2014. p. 3–29.

    Google Scholar 

  16. 16.

    Villaseñor A, Garcia-Perez I, Garcia A, Posma JM, Fernández-López M, Nicholas AJ, et al. Breast milk metabolome characterization in a single-phase extraction, multiplatform analytical approach. Anal Chem. 2014;86:8245–52.

    Article  Google Scholar 

  17. 17.

    Marincola FC, Noto A, Caboni P, Reali A, Barberini L, Lussu M, et al. A metabolomic study of preterm human and formula milk by high resolution NMR and GC/MS analysis: preliminary results. J Matern Fetal Neonatal Med. 2012;25:62–7.

    CAS  Article  Google Scholar 

  18. 18.

    Smilowitz JT, O’Sullivan A, Barile D, German JB, Lönnerdal B, Slupsky CM. The human milk metabolome reveals diverse oligosaccharide profiles. J Nutr. 2013;143:1709–18.

    CAS  Article  Google Scholar 

  19. 19.

    Praticò G, Capuani G, Tomassini A, Baldassarre ME, Delfini M, Miccheli A. Exploring human breast milk composition by NMR-based metabolomics. Nat Prod Res. 2014;28:95–101.

    Article  Google Scholar 

  20. 20.

    Dessì A, Briana D, Corbu S, Gavrili S, Marincola FC, Georgantzi S, et al. Metabolomics of breast milk: the importance of phenotypes. Metabolites. 2018;8.

  21. 21.

    Qian L, Zhao A, Zhang Y, Chen T, Zeisel SH, Jia W, et al. Metabolomic approaches to explore chemical diversity of human breast-milk, formula milk and bovine milk. Int J Mol Sci. 2016;17.

  22. 22.

    Garwolińska D, Hewelt-Belka W, Kot-Wasik A, Sundekilde UK. Nuclear magnetic resonance metabolomics reveals qualitative and quantitative differences in the composition of human breast milk and milk formulas. Nutrients. 2020;12:921.

    Article  Google Scholar 

  23. 23.

    Meoni G, Tenori L, Luchinat C. Nuclear magnetic resonance-based metabolomic comparison of breast milk and organic and traditional formula milk brands for infants and toddlers. OMICS. 2020;24:424–36.

    CAS  Article  Google Scholar 

  24. 24.

    Scano P, Murgia A, Demuru M, Consonni R, Caboni P. Metabolite profiles of formula milk compared to breast milk. Food Res Int. 2016;87:76–82.

    CAS  Article  Google Scholar 

  25. 25.

    Phan M, Momin SR, Senn MK, Wood AC. Metabolomic insights into the effects of breast milk versus formula milk feeding in infants. Curr Nutr Rep. 2019;8:295–306.

    Article  PubMed  Google Scholar 

  26. 26.

    Demmelmair H, Koletzko B. Variation of metabolite and hormone contents in human milk. Clin Perinatol. 2017;44:151–64.

    Article  Google Scholar 

  27. 27.

    Li K, Jiang J, Xiao H, Wu K, Qi C, Sun J, et al. Changes in the metabolite profile of breast milk over lactation stages and their relationship with dietary intake in Chinese women: HPLC-QTOFMS based metabolomic analysis. Food Funct. 2018;9:5189–97.

    CAS  Article  Google Scholar 

  28. 28.

    Wu J, Domellöf M, Zivkovic AM, Larsson G, Öhman A, Nording ML. NMR-based metabolite profiling of human milk: a pilot study of methods for investigating compositional changes during lactation. Biochem Biophys Res Commun. 2015;469:626–32.

    Article  Google Scholar 

  29. 29.

    Alexandre-Gouabau MC, Moyon T, David-Sochard A, Fenaille F, Cholet S, Royer AL, et al. Comprehensive preterm breast milk metabotype associated with optimal infant early growth pattern. Nutrients. 2019;11.

  30. 30.

    Longini M, Tataranno ML, Proietti F, Tortoriello M, Belvisi E, Vivi A, et al. A metabolomic study of preterm and term human and formula milk by proton MRS analysis: preliminary results. J Matern Neonatal Med. 2014;27:27–33.

    CAS  Article  Google Scholar 

  31. 31.

    Perrone S, Longini M, Zollino I, Bazzini F, Tassini M, Vivi A, et al. Breast milk: to each his own. From metabolomic study, evidence of personalized nutrition in preterm infants. Nutrition. 2019;62:158–61.

    Article  Google Scholar 

  32. 32.

    • Gay MCL, Koleva PT, Slupsky CM, et al. Worldwide variation in human milk metabolome: indicators of breast physiology and maternal lifestyle? Nutrients. 2018. metabolomic study on human milk from five different geographical origin pointing towards environment-related impact on the metabolome.

  33. 33.

    Gómez-Gallego C, Morales JM, Monleón D, du Toit E, Kumar H, Linderborg K, et al. Human breast milk NMR metabolomic profile across specific geographical locations and its association with the milk microbiota. Nutrients. 2018;10.

  34. 34.

    Bravi F, Wiens F, Decarli A, Pont AD, Agostoni C, Ferraroni M. Impact of maternal nutrition on breast-milk composition—a systematic review. Am J Clin Nutr. 2016;104:646–62.

    CAS  Article  Google Scholar 

  35. 35.

    Dangat K, Upadhyay D, Kilari A, Sharma U, Kemse N, Mehendale S, et al. Altered breast milk components in preeclampsia; an in-vitro proton NMR spectroscopy study. Clin Chim Acta. 2016;463:75–83.

    CAS  Article  Google Scholar 

  36. 36.

    Meng X, Dunsmore G, Koleva P, Elloumi Y, Wu RY, Sutton RT, et al. The profile of human milk metabolome, cytokines, and antibodies in inflammatory bowel diseases versus healthy mothers, and potential impact on the newborn. J Crohn's Colitis. 2019;13:431–41.

    Article  Google Scholar 

  37. 37.

    Wen L, Wu Y, Yang Y, Han TL, Wang W, Fu H, et al. Gestational diabetes mellitus changes the metabolomes of human colostrum, transition milk and mature milk. Med Sci Monit. 2019;25:6128–52.

    CAS  Article  Google Scholar 

  38. 38.

    Briana DD, Fotakis C, Kontogeorgou A, Gavrili S, Georgatzi S, Zoumpoulakis P, et al. Early human-milk metabolome in cases of intrauterine growth–restricted and macrosomic infants. J Parenter Enter Nutr. 2020;44:1510–8.

    CAS  Article  Google Scholar 

  39. 39.

    Carducci B, Zulfiqar AB. Care of the growth-restricted newborn. Best Pract Res Clin Obstet Gynaecol. 2018;49:103–16.

    Article  PubMed  Google Scholar 

  40. 40.

    • Saben JL, Sims CR, Piccolo BD, Andres A. Maternal adiposity alters the human milk metabolome: associations between nonglucose monosaccharides and infant adiposity. Am J Clin Nutr. 2020. investigation comprising a large cohort and sampling longitudinally over 6 months’ lactation. Including anthropometric measures from both the maternal part and related infant. Comprehensive analysis of the human milk metabolome.

  41. 41.

    • Bardanzellu F, Puddu M, Peroni DG, Fanos V. The human breast milk metabolome in overweight and obese mothers. Front Immunol. 2020;11:1533 Recent review on human milk metabolome and associations to maternal weight status. Primarily stressing that some evidence thus far points towards associations between maternal BMI and human milk metabolome, although comprehensive analyses further taking into account maternal genetics, diet etc. are necessary, along with studies investigating the potential impact in offspring.

    CAS  Article  Google Scholar 

  42. 42.

    Piñeiro-Ramos JD, Parra-Llorca A, Ten-Doménech I, et al. Effect of donor human milk on host-gut microbiota and metabolic interactions in preterm infants. Clin Nutr, in press. 2020.

Download references


The authors wish to thank the Sino-Danish Center for Education and Research for funding.

Author information




KOP: Conceptualization, literature search, writing of original draft and editing. UKS: Conceptualization, literature search, validation, editing and supervision. All authors approved the final version of the manuscript.

Corresponding author

Correspondence to Ulrik Kræmer Sundekilde.

Ethics declarations

Conflict of Interest

The authors declare they have no conflict of interest.

Human and Animal Rights and Informed Consent

This article does not contain any studies with human or animal subjects performed by any of the authors.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

This article is part of the Topical Collection on Maternal and Childhood Nutrition

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Poulsen, K.O., Sundekilde, U.K. The Metabolomic Analysis of Human Milk Offers Unique Insights into Potential Child Health Benefits. Curr Nutr Rep (2021).

Download citation


  • Human milk
  • Metabolomics
  • Infant metabolism
  • Infant nutrition