Determination of 2′-Fucosyllactose and Lacto-N-neotetraose in Infant Formula
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-neote...
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| Format: | Article |
| Language: | English |
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MDPI AG
2018-10-01
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| Series: | Molecules |
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| Online Access: | http://www.mdpi.com/1420-3049/23/10/2650 |
| _version_ | 1811274564748967936 |
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| author | Sean Austin Denis Cuany Julien Michaud Bernd Diehl Begoña Casado |
| author_facet | Sean Austin Denis Cuany Julien Michaud Bernd Diehl Begoña Casado |
| author_sort | Sean Austin |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T23:21:13Z |
| format | Article |
| id | doaj.art-3ae07a9cef8844ee99990d5a63f7ea95 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-04-12T23:21:13Z |
| publishDate | 2018-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-3ae07a9cef8844ee99990d5a63f7ea952022-12-22T03:12:31ZengMDPI AGMolecules1420-30492018-10-012310265010.3390/molecules23102650molecules23102650Determination of 2′-Fucosyllactose and Lacto-N-neotetraose in Infant FormulaSean Austin0Denis Cuany1Julien Michaud2Bernd Diehl3Begoña Casado4Nestlé Research, Vers-Chez-Les-Blanc, 1000 Lausanne, SwitzerlandNestlé Research, Vers-Chez-Les-Blanc, 1000 Lausanne, SwitzerlandNestlé Research, Vers-Chez-Les-Blanc, 1000 Lausanne, SwitzerlandSpectral Services, Emil-Hoffmann Strasse 33, D-50996 Köln, GermanyNestlé Research, Vers-Chez-Les-Blanc, 1000 Lausanne, SwitzerlandHuman 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.http://www.mdpi.com/1420-3049/23/10/2650human milk oligosaccharidesliquid chromatographyinfant formula2′-FLLNnT |
| spellingShingle | Sean Austin Denis Cuany Julien Michaud Bernd Diehl Begoña Casado Determination of 2′-Fucosyllactose and Lacto-N-neotetraose in Infant Formula Molecules human milk oligosaccharides liquid chromatography infant formula 2′-FL LNnT |
| title | Determination of 2′-Fucosyllactose and Lacto-N-neotetraose in Infant Formula |
| title_full | Determination of 2′-Fucosyllactose and Lacto-N-neotetraose in Infant Formula |
| title_fullStr | Determination of 2′-Fucosyllactose and Lacto-N-neotetraose in Infant Formula |
| title_full_unstemmed | Determination of 2′-Fucosyllactose and Lacto-N-neotetraose in Infant Formula |
| title_short | Determination of 2′-Fucosyllactose and Lacto-N-neotetraose in Infant Formula |
| title_sort | determination of 2 fucosyllactose and lacto n neotetraose in infant formula |
| topic | human milk oligosaccharides liquid chromatography infant formula 2′-FL LNnT |
| url | http://www.mdpi.com/1420-3049/23/10/2650 |
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