A nine-country study of the protein content and amino acid composition of mature human milk

  • Ping Feng Wyeth Nutrition
  • Ming Gao Wyeth Nutrition
  • Anita Burgher Wyeth Nutrition
  • Tian Hui Zhou Bio Tx Clinical Research, Pfizer Inc
  • Kathryn Pramuk Wyeth Nutrition
Keywords: Human milk, amino acid, protein, nitrogen, lactation stage


Background: Numerous studies have evaluated protein and amino acid levels in human milk. However, research in this area has been limited by small sample sizes and study populations with little ethnic or racial diversity.

Objective: Evaluate the protein and amino acid composition of mature (≥30 days) human milk samples collected from a large, multinational study using highly standardized methods for sample collection, storage, and analysis.

Design: Using a single, centralized laboratory, human milk samples from 220 women (30–188 days postpartum) from nine countries were analyzed for amino acid composition using Waters AccQ-Tag high-performance liquid chromatography and total nitrogen content using the LECO FP-528 nitrogen analyzer. Total protein was calculated as total nitrogen×6.25. True protein, which includes protein, free amino acids, and peptides, was calculated from the total amino acids.

Results: Mean total protein from individual countries (standard deviation [SD]) ranged from 1,133 (125.5) to 1,366 (341.4) mg/dL; the mean across all countries (SD) was 1,192 (200.9) mg/dL. Total protein, true protein, and amino acid composition were not significantly different across countries except Chile, which had higher total and true protein. Amino acid profiles (percent of total amino acids) did not differ across countries. Total and true protein concentrations and 16 of 18 amino acid concentrations declined with the stage of lactation.

Conclusions: Total protein, true protein, and individual amino acid concentrations in human milk steadily decline from 30 to 151 days of lactation, and are significantly higher in the second month of lactation compared with the following 4 months. There is a high level of consistency in the protein content and amino acid composition of human milk across geographic locations. The size and diversity of the study population and highly standardized procedures for the collection, storage, and analysis of human milk support the validity and broad application of these findings.

Keywords: human milk; amino acid; protein; nitrogen; lactation stage

(Published: 26 August 2016)

Citation: Food & Nutrition Research 2016, 60: 31042 - http://dx.doi.org/10.3402/fnr.v60.31042


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Author Biographies

Ping Feng, Wyeth Nutrition
Technical Director- NPD
Ming Gao, Wyeth Nutrition

Analytical Chemistry

Anita Burgher, Wyeth Nutrition
Director, Analytical and Quality Services
Tian Hui Zhou, Bio Tx Clinical Research, Pfizer Inc
Kathryn Pramuk, Wyeth Nutrition
Principal Scientist
How to Cite
Feng P., Gao M., Burgher A., Zhou T. H., & Pramuk K. (2016). A nine-country study of the protein content and amino acid composition of mature human milk. Food & Nutrition Research, 60. https://doi.org/10.3402/fnr.v60.31042
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