Abstract

Human milk oligosaccharides (HMO) are the third most abundant solid component of human milk. It is likely that they are responsible for at least some of the benefits experienced by breast-fed infants. Until recently HMO were absent from infant formula, but 2′-fucosyllactose (2′-FL) and lacto-N-neoteraose (LNnT) have recently become available as ingredients. The development of formula containing these HMO and the quality control of such formula require suitable methods for the accurate determination of the HMO. We developed two different approaches for analysis of 2′-FL and LNnT in formula; high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and hydrophilic interaction liquid chromatography with fluorescence detection (HILIC-FLD). In lab trials using blank formula spiked with the two oligosaccharides, both approaches worked well with recoveries of 94–111% (HPAEC-PAD) and 94–104% (HILIC-FLD) and RSD (iR) of 2.1–7.9% (HPAEC-PAD) and 2.0–7.4% (HILIC-FLD). However, when applied to products produced in a pilot plant, the HPAEC-PAD approach sometimes delivered results below those expected from the addition rate of the ingredients. We hypothesize that the oligosaccharides interact with the formula matrix during the production process and, during sample preparation for HPAEC-PAD those interactions have not been broken. The conditions required for labeling the HMO for detection by the FLD apparently disrupt those interactions, and result in improved recoveries. It is likely that both analytical approaches are appropriate if a suitable extraction process is used to recover the HMO.

Highlights

  • Human milk oligosaccharides (HMO) are the third most abundant solid component of human milk after lactose and fats

  • In this paper we describe two chromatographic methods for the determination of 20 -fucsyllactose (20 -FL) and LNnT in infant formula

  • In order to determine the quantity of oligosaccharides added to the formula by chromatography it was necessary to obtain good quality, quantitative, analytical standards

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Summary

Introduction

Human milk oligosaccharides (HMO) are the third most abundant solid component of human milk after lactose and fats. HMO are nondigestible oligosaccharides and, among other things, are believed to be implicated in protecting the infant from infections, and in the establishment of a healthy gut microbiota [1,2,3,4,5]. More than 150 different HMO have been identified [6], quantitative data only exists for about 30 of the HMO in milk [7]. The HMO can be roughly classified in to three different types; fucosylated HMO, sialylated. This is not a rigid separation of structures, since certain HMO can be both fucosylated and sialylated and can be classified in to two of the groups.

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