Application of a sensitive fluorometric HPLC assay to determine the sialic acid content of infant formulas

[EN] The developing human brain requires high amounts of sialic acids. While human milk is very rich in sialic acids, cow's milk based infant formulas provide lower amounts of sialic acids, and sialic acids are absent in soy milk based formulas. This has prompted interest in the supplementation...

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Autores: Martín Martín, María Jesús, Vázquez, Enrique, Rueda, Ricardo
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2007
País:España
Recursos:Universidad de Salamanca (USAL)
Repositorio:GREDOS. Repositorio Institucional de la Universidad de Salamanca
OAI Identifier:oai:gredos.usal.es:10366/167628
Acesso em linha:http://hdl.handle.net/10366/167628
Access Level:acceso embargado
Palavra-chave:Sialic acids
Infant formulas
HPLC
Ácidos siálicos
fórmulas infantiles
Infant Formula
Sialic Acids
2302 Bioquímica
ácidos siálicos
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spelling Application of a sensitive fluorometric HPLC assay to determine the sialic acid content of infant formulasMartín Martín, María JesúsVázquez, EnriqueRueda, RicardoSialic acidsInfant formulasHPLCÁcidos siálicosfórmulas infantilesInfant FormulaSialic Acids2302 Bioquímicaácidos siálicosfórmulas infantiles[EN] The developing human brain requires high amounts of sialic acids. While human milk is very rich in sialic acids, cow's milk based infant formulas provide lower amounts of sialic acids, and sialic acids are absent in soy milk based formulas. This has prompted interest in the supplementation of formulas with sialic acids, either free or bound to glycoconjugates. In order for fortification of infant formulas with sialic acids to be appropriate for the developmental needs of the infant, an accurate quantitation of sialic acid content of infant formulas through a reliable and easy-to-use method is, therefore, of great interest to industry. In the present method, we describe the application of one of the most widely used analytical techniques to the quantitation of sialic acids in infant formulas. Briefly, sialic acids are hydrolyzed from glycoconjugates, derivatized using 1,2-diamino-4,5-methylenedioxybenzene dihydrochloride (DMB), and separated using reversed-phase high-performance liquid chromatography. The method fulfilled the established criteria for validation, with an interassay standard deviation of less than 5%, accuracy greater than 97%, and surrogate recovery between 98 and 104%. An investigation of the ruggedness of the method identified two key criteria: both standards and samples must be subjected to the same temperature and pH conditions for an accurate quantitation; and prolonged storage (more than 2 days) of the DMB reagent and derivatives must be avoided. In conclusion, this method is specific, straightforward, and accurate and can be easily performed in a quality-assurance laboratory to track the level of sialic acid in formulas that contain both inherent and fortified amounts of sialic acids. Figure Infant formula and HPLC vials used for the sialic acid quantitation.Acknowledgements This work was partially supported by the Programa Torres Quevedo, from the Spanish Ministry of Education and Science and the European Social Fund. The authors would like to thank M.H. Dohnalek and P.A. Prieto for the critical review of the manuscript. Financiadores Ministerio de Ciencia e Innovación Spain European Social Fund European UnionAbbott LaboratoriesSpringer Natureinfo202520252007info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10366/167628reponame:GREDOS. Repositorio Institucional de la Universidad de Salamancainstname:Universidad de Salamanca (USAL)InglésAttribution-NonCommercial-NoDerivatives 4.0 Internacionalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/embargoedAccessoai:gredos.usal.es:10366/1676282026-06-07T06:28:51Z
dc.title.none.fl_str_mv Application of a sensitive fluorometric HPLC assay to determine the sialic acid content of infant formulas
title Application of a sensitive fluorometric HPLC assay to determine the sialic acid content of infant formulas
spellingShingle Application of a sensitive fluorometric HPLC assay to determine the sialic acid content of infant formulas
Martín Martín, María Jesús
Sialic acids
Infant formulas
HPLC
Ácidos siálicos
fórmulas infantiles
Infant Formula
Sialic Acids
2302 Bioquímica
ácidos siálicos
fórmulas infantiles
title_short Application of a sensitive fluorometric HPLC assay to determine the sialic acid content of infant formulas
title_full Application of a sensitive fluorometric HPLC assay to determine the sialic acid content of infant formulas
title_fullStr Application of a sensitive fluorometric HPLC assay to determine the sialic acid content of infant formulas
title_full_unstemmed Application of a sensitive fluorometric HPLC assay to determine the sialic acid content of infant formulas
title_sort Application of a sensitive fluorometric HPLC assay to determine the sialic acid content of infant formulas
dc.creator.none.fl_str_mv Martín Martín, María Jesús
Vázquez, Enrique
Rueda, Ricardo
author Martín Martín, María Jesús
author_facet Martín Martín, María Jesús
Vázquez, Enrique
Rueda, Ricardo
author_role author
author2 Vázquez, Enrique
Rueda, Ricardo
author2_role author
author
dc.subject.none.fl_str_mv Sialic acids
Infant formulas
HPLC
Ácidos siálicos
fórmulas infantiles
Infant Formula
Sialic Acids
2302 Bioquímica
ácidos siálicos
fórmulas infantiles
topic Sialic acids
Infant formulas
HPLC
Ácidos siálicos
fórmulas infantiles
Infant Formula
Sialic Acids
2302 Bioquímica
ácidos siálicos
fórmulas infantiles
description [EN] The developing human brain requires high amounts of sialic acids. While human milk is very rich in sialic acids, cow's milk based infant formulas provide lower amounts of sialic acids, and sialic acids are absent in soy milk based formulas. This has prompted interest in the supplementation of formulas with sialic acids, either free or bound to glycoconjugates. In order for fortification of infant formulas with sialic acids to be appropriate for the developmental needs of the infant, an accurate quantitation of sialic acid content of infant formulas through a reliable and easy-to-use method is, therefore, of great interest to industry. In the present method, we describe the application of one of the most widely used analytical techniques to the quantitation of sialic acids in infant formulas. Briefly, sialic acids are hydrolyzed from glycoconjugates, derivatized using 1,2-diamino-4,5-methylenedioxybenzene dihydrochloride (DMB), and separated using reversed-phase high-performance liquid chromatography. The method fulfilled the established criteria for validation, with an interassay standard deviation of less than 5%, accuracy greater than 97%, and surrogate recovery between 98 and 104%. An investigation of the ruggedness of the method identified two key criteria: both standards and samples must be subjected to the same temperature and pH conditions for an accurate quantitation; and prolonged storage (more than 2 days) of the DMB reagent and derivatives must be avoided. In conclusion, this method is specific, straightforward, and accurate and can be easily performed in a quality-assurance laboratory to track the level of sialic acid in formulas that contain both inherent and fortified amounts of sialic acids. Figure Infant formula and HPLC vials used for the sialic acid quantitation.
publishDate 2007
dc.date.none.fl_str_mv 2007
2025
2025
info
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10366/167628
url http://hdl.handle.net/10366/167628
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv Attribution-NonCommercial-NoDerivatives 4.0 Internacional
http://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/embargoedAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 Internacional
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv embargoedAccess
dc.publisher.none.fl_str_mv Springer Nature
publisher.none.fl_str_mv Springer Nature
dc.source.none.fl_str_mv reponame:GREDOS. Repositorio Institucional de la Universidad de Salamanca
instname:Universidad de Salamanca (USAL)
instname_str Universidad de Salamanca (USAL)
reponame_str GREDOS. Repositorio Institucional de la Universidad de Salamanca
collection GREDOS. Repositorio Institucional de la Universidad de Salamanca
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,811543

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