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 publicada
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.
Fil: Yonny, Melisa Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Reineri, Pablo Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Palma, Gustavo Adolfo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Tucumán-Santiago del Estero. Estación Experimental Agropecuaria Santiago del Estero; Argentina
Fil: Nazareno, Mónica Azucena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Materia
MALONDIALDEHYDE
SPERM CELLS
OXIDATIVE MARKER
BOVINE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/41576
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/41576
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semenYonny, Melisa EvangelinaReineri, Pablo SebastianPalma, Gustavo AdolfoNazareno, Mónica AzucenaMALONDIALDEHYDESPERM CELLSOXIDATIVE MARKERBOVINEhttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Frozen 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.Fil: Yonny, Melisa Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Reineri, Pablo Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaFil: Palma, Gustavo Adolfo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Tucumán-Santiago del Estero. Estación Experimental Agropecuaria Santiago del Estero; ArgentinaFil: Nazareno, Mónica Azucena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; ArgentinaRoyal Society of Chemistry2015-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/41576Yonny, Melisa Evangelina; Reineri, Pablo Sebastian; Palma, Gustavo Adolfo; Nazareno, Mónica Azucena; Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen; Royal Society of Chemistry; Analyst; 7; 15; 6-2015; 6331-63380003-2654CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1039/C5AY00950Binfo:eu-repo/semantics/altIdentifier/url/http://pubs.rsc.org/en/Content/ArticleLanding/2015/AY/C5AY00950Binfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:35:14Zoai:ri.conicet.gov.ar:11336/41576instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:35:14.328CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
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
MALONDIALDEHYDE
SPERM CELLS
OXIDATIVE MARKER
BOVINE
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 MALONDIALDEHYDE
SPERM CELLS
OXIDATIVE MARKER
BOVINE
topic MALONDIALDEHYDE
SPERM CELLS
OXIDATIVE MARKER
BOVINE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
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.
Fil: Yonny, Melisa Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Reineri, Pablo Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. Universidad Nacional de Santiago del Estero. Centro de Investigaciones y Transferencia de Santiago del Estero; Argentina
Fil: Palma, Gustavo Adolfo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Tucumán-Santiago del Estero. Estación Experimental Agropecuaria Santiago del Estero; Argentina
Fil: Nazareno, Mónica Azucena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Santiago del Estero. 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 2015-06
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/41576
Yonny, Melisa Evangelina; Reineri, Pablo Sebastian; Palma, Gustavo Adolfo; Nazareno, Mónica Azucena; Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen; Royal Society of Chemistry; Analyst; 7; 15; 6-2015; 6331-6338
0003-2654
CONICET Digital
CONICET
url http://hdl.handle.net/11336/41576
identifier_str_mv Yonny, Melisa Evangelina; Reineri, Pablo Sebastian; Palma, Gustavo Adolfo; Nazareno, Mónica Azucena; Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen; Royal Society of Chemistry; Analyst; 7; 15; 6-2015; 6331-6338
0003-2654
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1039/C5AY00950B
info:eu-repo/semantics/altIdentifier/url/http://pubs.rsc.org/en/Content/ArticleLanding/2015/AY/C5AY00950B
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
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 reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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