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. 2018 Oct 16;23(10):2650.
doi: 10.3390/molecules23102650.

Determination of 2'-Fucosyllactose and Lacto-N-neotetraose in Infant Formula

Sean Austin  1 , Denis Cuany  2 , Julien Michaud  3   4 , Bernd Diehl  5 , Begoña Casado  6   7
Affiliations

Determination of 2'-Fucosyllactose and Lacto-N-neotetraose in Infant Formula

Sean Austin et al. Molecules. .

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.

Keywords: 2′-FL; LNnT; human milk oligosaccharides; infant formula; liquid chromatography.

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Conflict of interest statement

S.A., J.M., D.C., and B.C. were all employees of Nestec Ltd. when this work was performed; B.D. declares no conflict of interest.

Figures

Figure 1
Figure 1
Separation of 2′-fucosyllactose (2′-FL) and Lacto-N-neotetraose (LNnT) from each other and from lactose in infant formula by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD).
Figure 2
Figure 2
Separation of 2′-fucosyllactose (2′-FL) and Lacto-N-neotetraose (LNnT) from each other and from lactose in infant formula by HILIC-FLD, using laminaritriose as an internal standard (Int. Std.).
See this image and copyright information in PMC

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