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Determination of Human Milk Oligosaccharides (HMO)

Detection of 2’-Fucosyllactose (2’FL) and Lacto-N-neotetraose (LNnT) in infant food

Dec 2021. The Eurofins Food & Feed Testing laboratory network in Germany now offers the determination of the oligosaccharides 2’-Fucosyllactose (2’FL) and Lacto-N-neotetraose (LNnT). The analysis is carried out using HPAEC-PAD and is accredited for infant formula and follow-on formula as well as food for special medical purposes (FSMP). The limits of quantification (LOQ) are 10 mg/100 g for solid and 4 mg/100 g for liquid products.

What are HMO? Definition of terms, occurrence and function

In terms of quantity, human milk oligosaccharides (HMO) can be found directly after milk sugar (lactose) and milk fats in human breast milk. They are increasingly becoming a focus of current research as they are believed to have beneficial effects on health and development in infants. There are more than 200 different HMOs varying in their structure. The most important so far include 2’-Fucosyllactose (2’FL) and Lacto-N-neotetraose (LNnT). They are based on lactose, are modified by the addition of e.g. sugars and form complex structures. Since recently, single HMOs can be technologically produced on a large scale and are thus available for food production.

Various effects are assigned to HMO^[1,2,3]:

Antimicrobial effect
If an HMO is built into the cell membrane of bacteria, they are no longer able to multiply^[4]. In addition, they can also reduce fungal attack by curbing hyphae growth^[5].

Anti-adhesive properties
Human intestinal epithelia have sugar molecules on their surface to which pathogenic germs dock. The structure of the HMO is similar to the sugar structures on the surface of the epithelial cells. The pathogens attach themselves to the unbound HMO and are channeled through the intestine and excreted.

Immune modulating effects
Part of the HMO is absorbed in the intestine and thus gets into the blood. The sugars have a positive effect on epithelial and immune cells throughout the body and support the defense against bacteria, e.g. in cystitis.

Prebiotic effect
In contrast to probiotics, which contain viable microorganisms such as lactic acid bacteria or yeast, prebiotics are indigestible food components (mostly based on carbohydrates) that promote the growth and activity of the microbiome. Prebiotics serve intestinal bacteria (e.g. bifidobacteria^[6]) as a food basis and can thus have a positive effect on the composition of the intestinal flora, which has a positive effect on human health.

Difference to galacto- and fructooligosaccharides (GOS and FOS/inulin)

FOS and GOS are also complex carbohydrates and belong to the group of prebiotics as well. However, GOS / FOS and HMO differ in their chemical structure: HMO show more complex structures with branched side chains, while GOS / FOS exist as linear, unbranched structures. GOS and FOS can also be analysed and determined in the Eurofins Food & Feed Testing laboratory network in Germany.

Legal background

The two HMO 2’-Fucosyllactose (2’FL) and Lacto-N-neotetraose (LNnT) have been classified as safe for food fortification by the European Food Safety Authority (EFSA) and the American Food and Drug Administration (FDA). According to the Commission's Implementing Decision (EU) 2016/376 of 11 March 2016^[7], 2'FL may be used in infant formula and follow-on formula in a maximum amount of 1.2 g / l alone or together with 0.6 g / l LNnT in a ratio of 2:1 and be placed on the EU market as a novel food. According to the Implementing Decision (EU) 2016/375^[8], the addition of LNnT in the maximum amount of 0.6 g / l is only permitted together with 2'FL in a ratio of 1:2.

Use of HMO

Infants who cannot be breastfed can also benefit from HMO, as these carbohydrates have recently been used in food for babies and toddlers as well as in foods for special medical purposes. We can now, as one of the first providers on the market, determine both HMOs and their legally prescribed ratio.

Have we piqued your interest?

If you would like to know more about the analysis of human milk oligosaccharides (HMO) or if you have any questions about our analysis, please do not hesitate to contact your personal account manager or our expert on the topic, Oliver Schwöbel.

Sources, relevant Implementing Decisions

[1] Bode L. Human milk oligosaccharides: Every baby needs a sugar mama. Glycobiology; 22: 1147–1162 (2012)
[2] Davis EC, Dinsmoor AM, Wang M, Donovan SM. Microbiome Composition in Pediatric Populations from Birth to Adolescence: Impact of Diet and Prebiotic and Probiotic Interventions. Dig Dis Sci 65: 706–722 (2020)
[3] Kunz C, Rudloff S. Health promoting aspects of milk oligosaccharides. Int Dairy J 16: 1341–1346 (2006)
[4] Bode L. The functional biology of human milk oligosaccharides. Early Hum Dev; 91: 619-622 (2015)
[5] Morozov V, Hansman G, Hanisch F-G et al. Human milk oligosaccharides as promising antivirals. Mol Nutr Food Res 2018; 62: 1700679 (2018)
[6] Thomson P, Medina D, Garrido D. Human milk oligosaccharides and infant gut bifidobacteria: Molecular strategies for their utilization. Food Microbiol; 75: 37–46 (2018)
[7] Commission Implementing Decision (EU) 2016/375 authorising the placing on the market of lacto-N-neotetraose as a novel food ingredient
[8] Commission Implementing Decision (EU) 2016/376 authorising the placing on the market of 2′-O-fucosyllactose as a novel food ingredient