Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen

Autores
Yonny, Melisa Evangelina; Reineri, Pablo Sebastian; Palma, Gustavo Adolfo; Nazareno, Mónica Azucena
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión aceptada
Descripción
Frozen sperm is widely used in artificial insemination of cattle as well as other animal species. As a consequence of the freezing and thawing processes of semen, an excess of reactive oxygen species (ROS) are formed. ROS produce oxidative damage in sperm cells affecting both motility and fertility. Malondialdehyde (MDA) is one of the most recognized biomarkers of an advanced oxidative status. MDA was analyzed after its condensation reaction with thiobarbituric acid (TBA); however, other molecules can also react with TBA. In order to determine specifically the MDA–TBA2 condensation product in cryopreserved bovine semen, a sensitive and selective separation strategy was developed using high performance liquid chromatography (HPLC) with diode array detection (DAD). This is the first report on MDA determination in bovine semen by a separation method. Different methodological approaches were assayed. Treatment A directly measured total MDA through acidic hydrolysis combined with TBA condensation in a single step. Treatment B evaluated separately the TBA condensation product of free MDA and protein bound MDA after its release with alkaline hydrolysis. The highest concentration of MDA was detected following treatment A. An HPLC method was developed and validated by comparing with the traditional spectrophotometric method. The detection and quantification limits were 0.034 μM and 0.086 μM. The DAD response was linear in the range between 0.086 and 9.1 μM. The recovery was 91%. The intra- and inter-day relative standard deviations were 3.7 and 3.8%, respectively. The proposed HPLC method was markedly more sensitive and more specific than the traditional spectrophotometric one.
EEA Santiago del Estero
Fil: Yonny, Melisa Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Yonny, Melisa Evangelina. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Reineri, Pablo Sebastian. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santiago del Estero; Argentina.
Fil: Palma, Gustavo Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Palma, Gustavo Adolfo. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Nazareno, Mónica Azucena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Nazareno, Mónica Azucena. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fuente
Analytical Methods 7 (15) : 1-33 (2015)
Materia
Ganado Bovino
Técnicas Analíticas
Criopreservación
Malondialdehído
Cattle
Semen
Analytical Methods
Cryopreservation
Malondialdehyde
Métodos Analíticos
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/14479
Acceder
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oai_identifier_str oai:localhost:20.500.12123/14479
network_acronym_str INTADig
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network_name_str INTA Digital (INTA)
spelling Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semenYonny, Melisa EvangelinaReineri, Pablo SebastianPalma, Gustavo AdolfoNazareno, Mónica AzucenaGanado BovinoTécnicas AnalíticasCriopreservaciónMalondialdehídoCattleSemenAnalytical MethodsCryopreservationMalondialdehydeMétodos AnalíticosFrozen sperm is widely used in artificial insemination of cattle as well as other animal species. As a consequence of the freezing and thawing processes of semen, an excess of reactive oxygen species (ROS) are formed. ROS produce oxidative damage in sperm cells affecting both motility and fertility. Malondialdehyde (MDA) is one of the most recognized biomarkers of an advanced oxidative status. MDA was analyzed after its condensation reaction with thiobarbituric acid (TBA); however, other molecules can also react with TBA. In order to determine specifically the MDA–TBA2 condensation product in cryopreserved bovine semen, a sensitive and selective separation strategy was developed using high performance liquid chromatography (HPLC) with diode array detection (DAD). This is the first report on MDA determination in bovine semen by a separation method. Different methodological approaches were assayed. Treatment A directly measured total MDA through acidic hydrolysis combined with TBA condensation in a single step. Treatment B evaluated separately the TBA condensation product of free MDA and protein bound MDA after its release with alkaline hydrolysis. The highest concentration of MDA was detected following treatment A. An HPLC method was developed and validated by comparing with the traditional spectrophotometric method. The detection and quantification limits were 0.034 μM and 0.086 μM. The DAD response was linear in the range between 0.086 and 9.1 μM. The recovery was 91%. The intra- and inter-day relative standard deviations were 3.7 and 3.8%, respectively. The proposed HPLC method was markedly more sensitive and more specific than the traditional spectrophotometric one.EEA Santiago del EsteroFil: Yonny, Melisa Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Yonny, Melisa Evangelina. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Reineri, Pablo Sebastian. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santiago del Estero; Argentina.Fil: Palma, Gustavo Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Palma, Gustavo Adolfo. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Nazareno, Mónica Azucena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Nazareno, Mónica Azucena. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaRoyal Society of Chemistry2023-04-14T12:53:50Z2023-04-14T12:53:50Z2015info:eu-repo/semantics/articleinfo:eu-repo/semantics/acceptedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/14479https://pubs.rsc.org/en/content/articlelanding/2015/ay/c5ay00950b1759-96601759-9679https://doi.org/10.1039/C5AY00950BAnalytical Methods 7 (15) : 1-33 (2015)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-10-16T09:31:09Zoai:localhost:20.500.12123/14479instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-10-16 09:31:09.902INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen
title Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen
spellingShingle Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen
Yonny, Melisa Evangelina
Ganado Bovino
Técnicas Analíticas
Criopreservación
Malondialdehído
Cattle
Semen
Analytical Methods
Cryopreservation
Malondialdehyde
Métodos Analíticos
title_short Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen
title_full Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen
title_fullStr Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen
title_full_unstemmed Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen
title_sort Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen
dc.creator.none.fl_str_mv Yonny, Melisa Evangelina
Reineri, Pablo Sebastian
Palma, Gustavo Adolfo
Nazareno, Mónica Azucena
author Yonny, Melisa Evangelina
author_facet Yonny, Melisa Evangelina
Reineri, Pablo Sebastian
Palma, Gustavo Adolfo
Nazareno, Mónica Azucena
author_role author
author2 Reineri, Pablo Sebastian
Palma, Gustavo Adolfo
Nazareno, Mónica Azucena
author2_role author
author
author
dc.subject.none.fl_str_mv Ganado Bovino
Técnicas Analíticas
Criopreservación
Malondialdehído
Cattle
Semen
Analytical Methods
Cryopreservation
Malondialdehyde
Métodos Analíticos
topic Ganado Bovino
Técnicas Analíticas
Criopreservación
Malondialdehído
Cattle
Semen
Analytical Methods
Cryopreservation
Malondialdehyde
Métodos Analíticos
dc.description.none.fl_txt_mv Frozen sperm is widely used in artificial insemination of cattle as well as other animal species. As a consequence of the freezing and thawing processes of semen, an excess of reactive oxygen species (ROS) are formed. ROS produce oxidative damage in sperm cells affecting both motility and fertility. Malondialdehyde (MDA) is one of the most recognized biomarkers of an advanced oxidative status. MDA was analyzed after its condensation reaction with thiobarbituric acid (TBA); however, other molecules can also react with TBA. In order to determine specifically the MDA–TBA2 condensation product in cryopreserved bovine semen, a sensitive and selective separation strategy was developed using high performance liquid chromatography (HPLC) with diode array detection (DAD). This is the first report on MDA determination in bovine semen by a separation method. Different methodological approaches were assayed. Treatment A directly measured total MDA through acidic hydrolysis combined with TBA condensation in a single step. Treatment B evaluated separately the TBA condensation product of free MDA and protein bound MDA after its release with alkaline hydrolysis. The highest concentration of MDA was detected following treatment A. An HPLC method was developed and validated by comparing with the traditional spectrophotometric method. The detection and quantification limits were 0.034 μM and 0.086 μM. The DAD response was linear in the range between 0.086 and 9.1 μM. The recovery was 91%. The intra- and inter-day relative standard deviations were 3.7 and 3.8%, respectively. The proposed HPLC method was markedly more sensitive and more specific than the traditional spectrophotometric one.
EEA Santiago del Estero
Fil: Yonny, Melisa Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Yonny, Melisa Evangelina. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Reineri, Pablo Sebastian. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santiago del Estero; Argentina.
Fil: Palma, Gustavo Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Palma, Gustavo Adolfo. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Nazareno, Mónica Azucena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Nazareno, Mónica Azucena. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
description Frozen sperm is widely used in artificial insemination of cattle as well as other animal species. As a consequence of the freezing and thawing processes of semen, an excess of reactive oxygen species (ROS) are formed. ROS produce oxidative damage in sperm cells affecting both motility and fertility. Malondialdehyde (MDA) is one of the most recognized biomarkers of an advanced oxidative status. MDA was analyzed after its condensation reaction with thiobarbituric acid (TBA); however, other molecules can also react with TBA. In order to determine specifically the MDA–TBA2 condensation product in cryopreserved bovine semen, a sensitive and selective separation strategy was developed using high performance liquid chromatography (HPLC) with diode array detection (DAD). This is the first report on MDA determination in bovine semen by a separation method. Different methodological approaches were assayed. Treatment A directly measured total MDA through acidic hydrolysis combined with TBA condensation in a single step. Treatment B evaluated separately the TBA condensation product of free MDA and protein bound MDA after its release with alkaline hydrolysis. The highest concentration of MDA was detected following treatment A. An HPLC method was developed and validated by comparing with the traditional spectrophotometric method. The detection and quantification limits were 0.034 μM and 0.086 μM. The DAD response was linear in the range between 0.086 and 9.1 μM. The recovery was 91%. The intra- and inter-day relative standard deviations were 3.7 and 3.8%, respectively. The proposed HPLC method was markedly more sensitive and more specific than the traditional spectrophotometric one.
publishDate 2015
dc.date.none.fl_str_mv 2015
2023-04-14T12:53:50Z
2023-04-14T12:53:50Z
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/acceptedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/14479
https://pubs.rsc.org/en/content/articlelanding/2015/ay/c5ay00950b
1759-9660
1759-9679
https://doi.org/10.1039/C5AY00950B
url http://hdl.handle.net/20.500.12123/14479
https://pubs.rsc.org/en/content/articlelanding/2015/ay/c5ay00950b
https://doi.org/10.1039/C5AY00950B
identifier_str_mv 1759-9660
1759-9679
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Royal Society of Chemistry
publisher.none.fl_str_mv Royal Society of Chemistry
dc.source.none.fl_str_mv Analytical Methods 7 (15) : 1-33 (2015)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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