Study shows HMO levels in breast milk to aid next-gen formula development

By Stephen Daniells

- Last updated on GMT

Human milk oligosaccharides (HMOs), unique carbohydrates that make up about 10% of the dry weight of mother’s milk, are beginning to be used in infant formula.   Image © Tom Merton / Getty Images
Human milk oligosaccharides (HMOs), unique carbohydrates that make up about 10% of the dry weight of mother’s milk, are beginning to be used in infant formula. Image © Tom Merton / Getty Images

Related tags HMO HMOs Prebiotics

A systematic review of dozens of studies has revealed that levels of the five most prevalent HMOs in breast milk vary widely, and infant formula development should accommodate this variability.

The five most prevalent HMOs (human milk oligosaccharides) are 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), Lacto-N-tetraose (LNT), 3'-sialyllactose (3'-SL), and 6'-sialyllactose (6'-SL). 2’-FL is the most abundant and can be found in levels up to 10 g per liter of breast milk, according to data published in Food and Chemical Toxicology.

“Considering the overarching goal of matching infant formula composition as closely as possible to human milk, the “appropriate” supplementation levels should accommodate the variability of the HMO levels found in human milk,” ​wrote scientists from Spherix Consulting Group, Inc., the University of Kentucky, and Chr. Hansen.

“Moreover, supplementation above the mean levels in human milk is not only appropriate, but safe, as demonstrated in extensive toxicology, tolerance, and clinical studies that directly support the safety of the HMOs, as well as other similar prebiotic ingredients,” ​they added.


HMOs are unique carbohydrates that make up about 10% of the dry weight of mother’s milk. HMOs are not easily digested, so experts postulate that their purpose is to jump-start the infant’s microbiome.

There are over 150 different HMOs. Since 2’-FL is the most abundant it is the most studied to-date. The majority of the science to date has focused on infants, with a study published in Gut​ (He et al., 2016, Vol. 65, pp. 33–46) by scientists from Harvard Medical School indicating that 2’-FL could reduce the inflammatory response to pathogenic bacteria.

In addition, a 2017 paper in the Journal of Pediatric Gastroenterology and Nutrition​ (Puccio et al., Vol. 64, pp. 624–631) reported that infants fed formula with 2′-FL and lacto-N-neotetraose (LNnT), another HMO, had lower levels of bronchitis and required fewer antibiotics.

Results of a study published in the Journal of Nutrition​ in 2016 by scientists at Abbott Nutrition found that infants fed a formula with 2’-FL had levels of inflammatory cytokines similar to those observed for breast feeding, and significantly lower than those observed for infants fed a control formula containing no 2’-FL. 

Getting closer to nature

The new review was commissioned by Chr Hansen, with Jesper Sig Mathiasen, senior vice president, Chr. Hansen HMO, commenting: “Breastfeeding is the best way to ensure infant health and recommended by WHO. At Chr. Hansen, we further aim to support the healthy development of infants that cannot be breastfed by providing HMOs as an ingredient and blend for infant formula.

“The study presents important statistical data to help support the level of appropriate HMO supplementation in infant formula and confirms the safety of intake at concentrations higher than average. We see it as yet another testimony to our HMO offering,”​ he said.


The new data found that levels of the five most prevalent HMOs vary depending on the mother’s health and genetics, environmental and geographical factors, gestational age, and lactation stage.

The mean levels of the HMOs were found to be: 2.6 g/L for 2'-FL, 0.6 g/L for 3-FL,0.9 g/L for LNT, 0.3 g/L for 3'-SL, and 0.4 g/L for 6'-sialyllactose, with significant standard deviations for each showing the wide variability.

The data appears to validate the levels used in Chr Hansen’s 5HMO-Mix​, which is composed of: 2.99 g/L 2′-FL, 0.75 g/L 3-FL, 1.5 g/L LNT, 0.23 g/L 3′-SL, and 0.28 g/L 6′-SL

The product was used in a recent clinical study that supported the safety and tolerability of the HMO-enriched formula, with data published in Nutrients​ showing that the 5HMO-Mix and breastfed infants had softer stools at a higher stool frequency, compared to infants fed a control formula.

Chr Hansen’s Mathiasen commented: “We are excited to publish this study of the five most prevalent HMOs in breast milk, which are all included in Chr. Hansen’s 5HMO-Mix in concentrations closer to nature and respecting the natural variations.”

Source: Food and Chemical Toxicology
Volume 163, May 2022, 112877, doi: 10.1016/j.fct.2022.112877
“Weighted analysis of 2′-fucosyllactose, 3-fucosyllactose, lacto-N-tetraose, 3′-sialyllactose, and 6′-sialyllactose concentrations in human milk”
Authors: D.B. Conze et al.

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