Publication
Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography
| Summary: | The profiling analysis of catecholamines and their metabolites in brain tissue offers a crucial key to understand their functions in the body and the opportunity to follow up neural diseases. A rapid and simple liquid chromatography-fluorescence detection (LC-FLD) method was developed and validated for simultaneously measuring several catecholamines and endogenous related compounds in the rat brain tissue samples. The target analytes measured in this bioanalytical assay were levodopa (L-DOPA), dopamine (DA), norepinephrine (NE), epinephrine (E), 3-O-methyldopa (3-O-MD), and homovanillic acid (HVA), being the 3,4-dihydroxybenzylamine (DHBA) used as internal standard (IS). The six analytes (L-DOPA, DA, NE, E, 3-O-MD and HVA) can be determined in a single chromatographic run of less than 12min, and all the compounds (analytes and IS) were detected using their native fluorescence and monitored at excitation/emission wavelengths of 279nm/320nm, respectively. The chromatographic and detection conditions were experimentally optimized and then several validation parameters (linearity, limits of quantification and detection, precision and accuracy, recovery, stability and selectivity) were examined. In accordance with the international guidelines of the Food and Drug Administration and European Medicines Agency the method described herein exhibited limits of quantification in the range of 2-25ngmL-1, linearity in wide concentration ranges (r2≥0.994), and acceptable precision (coefficient variation ≤8.76%) and accuracy (bias ±14.65%) levels. Since the bioanalytical procedure does not involve pre-purification or derivatization of the sample, the absolute recovery was found to be around 100%. Moreover, the developed LC-FLD method was successfully applied for the determination of the compounds of interest in tissue samples of different rat brain regions (cerebellum, amygdala, cortex, hippocampus, striatum, mesencephalon, medulla oblongata, substantia nigra and ventral tegmental area). Hence, this assay represents a valuable bioanalytical tool to support several pre(non)clinical studies in the broad field of neurosciences, requiring the quantitative analysis of these bioamines and their metabolites. |
|---|---|
| Main Authors: | Fonseca, Beatriz |
| Other Authors: | Rodrigues, Márcio; Cristóvão, Ana; Gonçalves, Daniela; Fortuna, Ana; Bernardino, Liliana; Falcão, Amílcar; Alves, Gilberto |
| Subject: | Catecholamines Native fluorescence Brain tissue Bioanalytical method validation |
| Year: | 2017 |
| Country: | Portugal |
| Document type: | article |
| Access type: | open access |
| Associated institution: | Universidade Politécnica da Guarda |
| Language: | English |
| Origin: | Repositório Científico da Universidade Politécnica da Guarda |
| _version_ | 1880131245143228416 |
|---|---|
| author | Fonseca, Beatriz |
| author2 | Rodrigues, Márcio Cristóvão, Ana Gonçalves, Daniela Fortuna, Ana Bernardino, Liliana Falcão, Amílcar Alves, Gilberto |
| author2_role | author author author author author author author |
| author_facet | Fonseca, Beatriz Rodrigues, Márcio Cristóvão, Ana Gonçalves, Daniela Fortuna, Ana Bernardino, Liliana Falcão, Amílcar Alves, Gilberto |
| author_role | author |
| contributor_name_str_mv | Repositório Científico da Universidade Politécnica da Guarda |
| country_str | PT |
| creators_json_txt | [{\"Person.name\":\"Fonseca, Beatriz\"},{\"Person.name\":\"Rodrigues, Márcio\"},{\"Person.name\":\"Cristóvão, Ana\"},{\"Person.name\":\"Gonçalves, Daniela\"},{\"Person.name\":\"Fortuna, Ana\"},{\"Person.name\":\"Bernardino, Liliana\"},{\"Person.name\":\"Falcão, Amílcar\"},{\"Person.name\":\"Alves, Gilberto\"}] |
| datacite.contributors.contributor.contributorName.fl_str_mv | Repositório Científico da Universidade Politécnica da Guarda |
| datacite.creators.creator.creatorName.fl_str_mv | Fonseca, Beatriz Rodrigues, Márcio Cristóvão, Ana Gonçalves, Daniela Fortuna, Ana Bernardino, Liliana Falcão, Amílcar Alves, Gilberto |
| datacite.date.Accepted.fl_str_mv | 2017-04-01T00:00:00Z |
| datacite.rights.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| datacite.subjects.subject.fl_str_mv | Catecholamines Native fluorescence Brain tissue Bioanalytical method validation |
| datacite.titles.title.fl_str_mv | Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography |
| dc.contributor.none.fl_str_mv | Repositório Científico da Universidade Politécnica da Guarda |
| dc.creator.none.fl_str_mv | Fonseca, Beatriz Rodrigues, Márcio Cristóvão, Ana Gonçalves, Daniela Fortuna, Ana Bernardino, Liliana Falcão, Amílcar Alves, Gilberto |
| dc.date.Accepted.fl_str_mv | 2017-04-01T00:00:00Z |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | https://hdl.handle.net/10314/3607 |
| dc.language.none.fl_str_mv | eng |
| dc.publisher.none.fl_str_mv | Elsevier B.V |
| dc.rights.none.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| dc.subject.none.fl_str_mv | Catecholamines Native fluorescence Brain tissue Bioanalytical method validation |
| dc.title.fl_str_mv | Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography |
| dc.type.none.fl_str_mv | http://purl.org/coar/resource_type/c_6501 |
| description | The profiling analysis of catecholamines and their metabolites in brain tissue offers a crucial key to understand their functions in the body and the opportunity to follow up neural diseases. A rapid and simple liquid chromatography-fluorescence detection (LC-FLD) method was developed and validated for simultaneously measuring several catecholamines and endogenous related compounds in the rat brain tissue samples. The target analytes measured in this bioanalytical assay were levodopa (L-DOPA), dopamine (DA), norepinephrine (NE), epinephrine (E), 3-O-methyldopa (3-O-MD), and homovanillic acid (HVA), being the 3,4-dihydroxybenzylamine (DHBA) used as internal standard (IS). The six analytes (L-DOPA, DA, NE, E, 3-O-MD and HVA) can be determined in a single chromatographic run of less than 12min, and all the compounds (analytes and IS) were detected using their native fluorescence and monitored at excitation/emission wavelengths of 279nm/320nm, respectively. The chromatographic and detection conditions were experimentally optimized and then several validation parameters (linearity, limits of quantification and detection, precision and accuracy, recovery, stability and selectivity) were examined. In accordance with the international guidelines of the Food and Drug Administration and European Medicines Agency the method described herein exhibited limits of quantification in the range of 2-25ngmL-1, linearity in wide concentration ranges (r2≥0.994), and acceptable precision (coefficient variation ≤8.76%) and accuracy (bias ±14.65%) levels. Since the bioanalytical procedure does not involve pre-purification or derivatization of the sample, the absolute recovery was found to be around 100%. Moreover, the developed LC-FLD method was successfully applied for the determination of the compounds of interest in tissue samples of different rat brain regions (cerebellum, amygdala, cortex, hippocampus, striatum, mesencephalon, medulla oblongata, substantia nigra and ventral tegmental area). Hence, this assay represents a valuable bioanalytical tool to support several pre(non)clinical studies in the broad field of neurosciences, requiring the quantitative analysis of these bioamines and their metabolites. |
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| identifier.other.fl_str_mv | https://hdl.handle.net/10314/3607 |
| inst_facet_str | urn:organizationAcronym:upg{{{_:::_}}}Universidade Politécnica da Guarda |
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| institution | Universidade Politécnica da Guarda |
| instname_str | Universidade Politécnica da Guarda |
| language | eng |
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| organization_str_mv | urn:organizationAcronym:upg |
| person_str_mv | Fonseca, Beatriz Rodrigues, Márcio Cristóvão, Ana Gonçalves, Daniela Fortuna, Ana Bernardino, Liliana Falcão, Amílcar Alves, Gilberto |
| publishDate | 2017 |
| publisher.none.fl_str_mv | Elsevier B.V |
| repo_facet_str | urn:repositoryAcronym:ripg{{{_:::_}}}Repositório Científico da Universidade Politécnica da Guarda |
| reponame_str | Repositório Científico da Universidade Politécnica da Guarda |
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| service_str_mv | urn:repositoryAcronym:ripg |
| spelling | engElsevier B.VenThe profiling analysis of catecholamines and their metabolites in brain tissue offers a crucial key to understand their functions in the body and the opportunity to follow up neural diseases. A rapid and simple liquid chromatography-fluorescence detection (LC-FLD) method was developed and validated for simultaneously measuring several catecholamines and endogenous related compounds in the rat brain tissue samples. The target analytes measured in this bioanalytical assay were levodopa (L-DOPA), dopamine (DA), norepinephrine (NE), epinephrine (E), 3-O-methyldopa (3-O-MD), and homovanillic acid (HVA), being the 3,4-dihydroxybenzylamine (DHBA) used as internal standard (IS). The six analytes (L-DOPA, DA, NE, E, 3-O-MD and HVA) can be determined in a single chromatographic run of less than 12min, and all the compounds (analytes and IS) were detected using their native fluorescence and monitored at excitation/emission wavelengths of 279nm/320nm, respectively. The chromatographic and detection conditions were experimentally optimized and then several validation parameters (linearity, limits of quantification and detection, precision and accuracy, recovery, stability and selectivity) were examined. In accordance with the international guidelines of the Food and Drug Administration and European Medicines Agency the method described herein exhibited limits of quantification in the range of 2-25ngmL-1, linearity in wide concentration ranges (r2≥0.994), and acceptable precision (coefficient variation ≤8.76%) and accuracy (bias ±14.65%) levels. Since the bioanalytical procedure does not involve pre-purification or derivatization of the sample, the absolute recovery was found to be around 100%. Moreover, the developed LC-FLD method was successfully applied for the determination of the compounds of interest in tissue samples of different rat brain regions (cerebellum, amygdala, cortex, hippocampus, striatum, mesencephalon, medulla oblongata, substantia nigra and ventral tegmental area). Hence, this assay represents a valuable bioanalytical tool to support several pre(non)clinical studies in the broad field of neurosciences, requiring the quantitative analysis of these bioamines and their metabolites.application/pdfenDetermination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatographyFonseca, BeatrizRodrigues, MárcioCristóvão, AnaGonçalves, DanielaFortuna, AnaBernardino, LilianaFalcão, AmílcarAlves, GilbertoHostingInstitutionOrganizationalRepositório Científico da Universidade Politécnica da Guardae-mailmailto:no-reply+repositorio@ipg.ptno-reply+repositorio@ipg.pt2017-042017-04-01T00:00:00ZHANDLEhttps://hdl.handle.net/10314/3607http://purl.org/coar/access_right/c_abf2open accessCatecholaminesNative fluorescenceBrain tissueBioanalytical method validation225613 bytesliteraturehttp://purl.org/coar/resource_type/c_6501journal articlehttp://purl.org/coar/access_right/c_abf2application/pdfhttps://repositorio.ipg.pt/bitstreams/7faf7ddc-a395-4247-9180-cbf572772d82/download |
| spellingShingle | Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography Fonseca, Beatriz Catecholamines Native fluorescence Brain tissue Bioanalytical method validation |
| status | SINGLETON |
| subject.fl_str_mv | Catecholamines Native fluorescence Brain tissue Bioanalytical method validation |
| title | Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography |
| title_full | Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography |
| title_fullStr | Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography |
| title_full_unstemmed | Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography |
| title_short | Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography |
| title_sort | Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography |
| topic | Catecholamines Native fluorescence Brain tissue Bioanalytical method validation |
| topic_facet | Catecholamines Native fluorescence Brain tissue Bioanalytical method validation |
| url | https://hdl.handle.net/10314/3607 |
| visible | 1 |
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