HPLC–DAD Method for the Quantification of Carbamazepine, Oxcarbazepine and their Active Metabolites in HepaRG Cell Culture Samples
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10314/3102 https://doi.org/10.1007/s10337-016-3063-7 |
Resumo: | A new, sensitive and fast high-performance liquid chromatography–diode-array detection assay is herein reported, for the first time, to simultaneously quantify carbamazepine (CBZ), oxcarbazepine (OXC), and the active metabolites carbamazepine-10,11-epoxide (CBZ-E) and licarbazepine (LIC) in HepaRG cell culture medium samples. Chromatographic separation of analytes (CBZ, CBZE, OXC, LIC) and internal standard (IS) was achieved in less than 15 min on a C18-column, at 35 °C, using a mobile phase composed of water/methanol/acetonitrile (69:25:6 v/v/v) pumped at 1 mL min−1. The analytes and IS were detected at 215 nm. The method proved to be selective, accurate (bias ± 14.6 %), precise (coefficient of variation ≤13.1 %) and linear (r2 ≥ 0.9901) over the concentration ranges of 0.1–15 μg mL−1 for CBZ; 0.1–5 μg mL−1 for CBZ-E and OXC; and 0.1–40 μg mL−1 for LIC. Furthermore, the absolute recovery of the analytes ranged from 64.5 to 96.9 % and their stability was demonstrated in the studied conditions. This validated HPLC assay will be a suitable tool to support future in vitro metabolism profiling, drug interaction and other pharmacokinetic-based studies in HepaRG cells involving these antiepileptic drugs (CBZ and OXC) and their main metabolites. |
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HPLC–DAD Method for the Quantification of Carbamazepine, Oxcarbazepine and their Active Metabolites in HepaRG Cell Culture SamplesCarbamazepineOxcarbazepineHepaRG cellsHigh-performance liquid chromatographyIn vitro metabolism and drug interaction studiesA new, sensitive and fast high-performance liquid chromatography–diode-array detection assay is herein reported, for the first time, to simultaneously quantify carbamazepine (CBZ), oxcarbazepine (OXC), and the active metabolites carbamazepine-10,11-epoxide (CBZ-E) and licarbazepine (LIC) in HepaRG cell culture medium samples. Chromatographic separation of analytes (CBZ, CBZE, OXC, LIC) and internal standard (IS) was achieved in less than 15 min on a C18-column, at 35 °C, using a mobile phase composed of water/methanol/acetonitrile (69:25:6 v/v/v) pumped at 1 mL min−1. The analytes and IS were detected at 215 nm. The method proved to be selective, accurate (bias ± 14.6 %), precise (coefficient of variation ≤13.1 %) and linear (r2 ≥ 0.9901) over the concentration ranges of 0.1–15 μg mL−1 for CBZ; 0.1–5 μg mL−1 for CBZ-E and OXC; and 0.1–40 μg mL−1 for LIC. Furthermore, the absolute recovery of the analytes ranged from 64.5 to 96.9 % and their stability was demonstrated in the studied conditions. This validated HPLC assay will be a suitable tool to support future in vitro metabolism profiling, drug interaction and other pharmacokinetic-based studies in HepaRG cells involving these antiepileptic drugs (CBZ and OXC) and their main metabolites.Springer-Verlag Berlin Heidelberg2016-11-14T14:56:41Z2016-11-142016-03-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10314/3102http://hdl.handle.net/10314/3102https://doi.org/10.1007/s10337-016-3063-7engFerreira, AnaRodrigues, MárcioFalcão, AmílcarAlves, Gilbertoinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-01-14T02:56:23Zoai:bdigital.ipg.pt:10314/3102Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T01:42:31.209251Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
HPLC–DAD Method for the Quantification of Carbamazepine, Oxcarbazepine and their Active Metabolites in HepaRG Cell Culture Samples |
title |
HPLC–DAD Method for the Quantification of Carbamazepine, Oxcarbazepine and their Active Metabolites in HepaRG Cell Culture Samples |
spellingShingle |
HPLC–DAD Method for the Quantification of Carbamazepine, Oxcarbazepine and their Active Metabolites in HepaRG Cell Culture Samples Ferreira, Ana Carbamazepine Oxcarbazepine HepaRG cells High-performance liquid chromatography In vitro metabolism and drug interaction studies |
title_short |
HPLC–DAD Method for the Quantification of Carbamazepine, Oxcarbazepine and their Active Metabolites in HepaRG Cell Culture Samples |
title_full |
HPLC–DAD Method for the Quantification of Carbamazepine, Oxcarbazepine and their Active Metabolites in HepaRG Cell Culture Samples |
title_fullStr |
HPLC–DAD Method for the Quantification of Carbamazepine, Oxcarbazepine and their Active Metabolites in HepaRG Cell Culture Samples |
title_full_unstemmed |
HPLC–DAD Method for the Quantification of Carbamazepine, Oxcarbazepine and their Active Metabolites in HepaRG Cell Culture Samples |
title_sort |
HPLC–DAD Method for the Quantification of Carbamazepine, Oxcarbazepine and their Active Metabolites in HepaRG Cell Culture Samples |
author |
Ferreira, Ana |
author_facet |
Ferreira, Ana Rodrigues, Márcio Falcão, Amílcar Alves, Gilberto |
author_role |
author |
author2 |
Rodrigues, Márcio Falcão, Amílcar Alves, Gilberto |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Ferreira, Ana Rodrigues, Márcio Falcão, Amílcar Alves, Gilberto |
dc.subject.por.fl_str_mv |
Carbamazepine Oxcarbazepine HepaRG cells High-performance liquid chromatography In vitro metabolism and drug interaction studies |
topic |
Carbamazepine Oxcarbazepine HepaRG cells High-performance liquid chromatography In vitro metabolism and drug interaction studies |
description |
A new, sensitive and fast high-performance liquid chromatography–diode-array detection assay is herein reported, for the first time, to simultaneously quantify carbamazepine (CBZ), oxcarbazepine (OXC), and the active metabolites carbamazepine-10,11-epoxide (CBZ-E) and licarbazepine (LIC) in HepaRG cell culture medium samples. Chromatographic separation of analytes (CBZ, CBZE, OXC, LIC) and internal standard (IS) was achieved in less than 15 min on a C18-column, at 35 °C, using a mobile phase composed of water/methanol/acetonitrile (69:25:6 v/v/v) pumped at 1 mL min−1. The analytes and IS were detected at 215 nm. The method proved to be selective, accurate (bias ± 14.6 %), precise (coefficient of variation ≤13.1 %) and linear (r2 ≥ 0.9901) over the concentration ranges of 0.1–15 μg mL−1 for CBZ; 0.1–5 μg mL−1 for CBZ-E and OXC; and 0.1–40 μg mL−1 for LIC. Furthermore, the absolute recovery of the analytes ranged from 64.5 to 96.9 % and their stability was demonstrated in the studied conditions. This validated HPLC assay will be a suitable tool to support future in vitro metabolism profiling, drug interaction and other pharmacokinetic-based studies in HepaRG cells involving these antiepileptic drugs (CBZ and OXC) and their main metabolites. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-11-14T14:56:41Z 2016-11-14 2016-03-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10314/3102 http://hdl.handle.net/10314/3102 https://doi.org/10.1007/s10337-016-3063-7 |
url |
http://hdl.handle.net/10314/3102 https://doi.org/10.1007/s10337-016-3063-7 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Springer-Verlag Berlin Heidelberg |
publisher.none.fl_str_mv |
Springer-Verlag Berlin Heidelberg |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
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1799136918430023680 |