Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen
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....
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| Format: | Artículo |
| Language: | Inglés |
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Royal Society of Chemistry
2023
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| Online Access: | http://hdl.handle.net/20.500.12123/14479 https://pubs.rsc.org/en/content/articlelanding/2015/ay/c5ay00950b https://doi.org/10.1039/C5AY00950B |
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| author | Yonny, Melisa Evangelina Reineri, Pablo Sebastian Palma, Gustavo Adolfo Nazareno, Mónica Azucena |
| author_browse | Nazareno, Mónica Azucena Palma, Gustavo Adolfo Reineri, Pablo Sebastian Yonny, Melisa Evangelina |
| author_facet | Yonny, Melisa Evangelina Reineri, Pablo Sebastian Palma, Gustavo Adolfo Nazareno, Mónica Azucena |
| author_sort | Yonny, Melisa Evangelina |
| collection | INTA Digital |
| 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. |
| format | Artículo |
| id | INTA14479 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | Royal Society of Chemistry |
| publisherStr | Royal Society of Chemistry |
| record_format | dspace |
| spelling | INTA144792023-04-14T12:55:31Z Development of an analytical method to determine malondialdehyde as an oxidative marker in cryopreserved bovine semen Yonny, Melisa Evangelina Reineri, Pablo Sebastian Palma, Gustavo Adolfo Nazareno, Mónica Azucena Ganado Bovino Técnicas Analíticas Criopreservación Malondialdehído Cattle Semen Analytical Methods Cryopreservation Malondialdehyde Métodos Analíticos 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 2023-04-14T12:53:50Z 2023-04-14T12:53:50Z 2015 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion 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 eng 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) application/pdf Royal Society of Chemistry Analytical Methods 7 (15) : 1-33 (2015) |
| spellingShingle | Ganado Bovino Técnicas Analíticas Criopreservación Malondialdehído Cattle Semen Analytical Methods Cryopreservation Malondialdehyde Métodos Analíticos 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 |
| title | 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_short | 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 |
| topic | Ganado Bovino Técnicas Analíticas Criopreservación Malondialdehído Cattle Semen Analytical Methods Cryopreservation Malondialdehyde Métodos Analíticos |
| 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 |
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