Desenvolvimento e validação de metodologias para avaliação de ezetimiba por cromatografia líquida e espectrometria de massas
Autor(a) principal: | |
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Data de Publicação: | 2007 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/6052 |
Resumo: | Ezetimibe selectively inhibits the intestinal absorption of dietary cholesterol and related plant sterols, from the 2-azetidinones group, and is used for the treatment of hypercholesterolemia and phytosterolemia. The methodologies for the evaluation of ezetimibe in pharmaceutical products and plasma were developed and validated in the present work. The reversed-phase liquid chromatography (RP-LC) analysis was carried out using a Synergi fusion C18 column (150 mm x 4.6 mm), maintained at 45 oC. The mobile phase consisted of potassium phosphate buffer 0.03 M, pH 4.5/acetonitrile (35:65, V/V), run at a flow rate of 0.6 mL/min with detection at 234 nm. The chromatographic separation was obtained within 15 min and it was linear in the concentration range of 0.5-200 Hg/mL. The method was sucessfuly applied for the simultaneous determination of ezetimibe and simvastatin in pharmaceutical products. The liquid chromatography-tandem mass spectrometry (LCMS/ MS) method was developed and validated using a Luna C18 column (50 mm x 3.0 mm) and the mobile phase consisted of acetonitrile:water (85:15, V/V), run at a flow rate of 0.4 mL/min. The mass spectrometer, equipped with electrospray positive source, was used in multiple reaction monitoring mode (MRM), monitoring the transitions of 392.0>161.0 and 359.3>280.0, for ezetimibe and etoricoxib (internal standard), respectively. The chromatographic separation was obtained within 2 min and it was linear in the concentration range of 0.25-20 ng/mL (ezetimibe) and 1-300 ng/mL (ezetimibe and its glucuronide matabolite). The procedures were validated evaluating parameters such as the specificity, linearity, precision, accuracy, robustness, limit of detection and limit of quantitation. Besides, for the bioanalytical method, the matrix effects, recovery and stability studies were also analyzed, giving results within the acceptable range. The proposed method was applied for the analysis of pharmaceutical products, showing significant correlation (P>0.05) of the results. Moreover, the liquid-liquid extraction method developed and optimized allowed high mean recoveries of ezetimibe and internal standard from the plasma samples. The procedures can be applied for the biovailability studies and for the quality control of pharmaceutical products. |
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Desenvolvimento e validação de metodologias para avaliação de ezetimiba por cromatografia líquida e espectrometria de massasDevelopment and validation of methodologies for the evaluation of ezetimibe by liquid chromatography and mass spectrometryCromatografia líquidaEspectrometria de massasEzetimibaValidaçãoPlasma humanoProdutos farmacêuticosLiquid chromatographyMass spectrometryEzetimibeValidationHuman plasmaPharmaceutical productsCNPQ::CIENCIAS DA SAUDE::FARMACIAEzetimibe selectively inhibits the intestinal absorption of dietary cholesterol and related plant sterols, from the 2-azetidinones group, and is used for the treatment of hypercholesterolemia and phytosterolemia. The methodologies for the evaluation of ezetimibe in pharmaceutical products and plasma were developed and validated in the present work. The reversed-phase liquid chromatography (RP-LC) analysis was carried out using a Synergi fusion C18 column (150 mm x 4.6 mm), maintained at 45 oC. The mobile phase consisted of potassium phosphate buffer 0.03 M, pH 4.5/acetonitrile (35:65, V/V), run at a flow rate of 0.6 mL/min with detection at 234 nm. The chromatographic separation was obtained within 15 min and it was linear in the concentration range of 0.5-200 Hg/mL. The method was sucessfuly applied for the simultaneous determination of ezetimibe and simvastatin in pharmaceutical products. The liquid chromatography-tandem mass spectrometry (LCMS/ MS) method was developed and validated using a Luna C18 column (50 mm x 3.0 mm) and the mobile phase consisted of acetonitrile:water (85:15, V/V), run at a flow rate of 0.4 mL/min. The mass spectrometer, equipped with electrospray positive source, was used in multiple reaction monitoring mode (MRM), monitoring the transitions of 392.0>161.0 and 359.3>280.0, for ezetimibe and etoricoxib (internal standard), respectively. The chromatographic separation was obtained within 2 min and it was linear in the concentration range of 0.25-20 ng/mL (ezetimibe) and 1-300 ng/mL (ezetimibe and its glucuronide matabolite). The procedures were validated evaluating parameters such as the specificity, linearity, precision, accuracy, robustness, limit of detection and limit of quantitation. Besides, for the bioanalytical method, the matrix effects, recovery and stability studies were also analyzed, giving results within the acceptable range. The proposed method was applied for the analysis of pharmaceutical products, showing significant correlation (P>0.05) of the results. Moreover, the liquid-liquid extraction method developed and optimized allowed high mean recoveries of ezetimibe and internal standard from the plasma samples. The procedures can be applied for the biovailability studies and for the quality control of pharmaceutical products.Ezetimiba é um inibidor seletivo da absorção intestinal do colesterol e de fitosteróis, pertencente ao grupo das 2-ezetidinonas, indicado para o tratamento da hipercolesterolemia e fitosterolemia. No presente trabalho foram desenvolvidas e validadas metodologias para avaliação de ezetimiba em produtos farmacêuticos e plasma humano. As análises por cromatografia líquida em fase reversa (CL-FR) foram realizadas utilizando coluna Synergi fusion C18 (150 mm x 4,6 mm), mantida a 45 oC. A fase móvel foi composta de tampão fosfato de potássio 0,03 M, pH 4,5/acetonitrila (35:65, V/V), eluída na vazão de 0,6 mL/min e detecção no ultravioleta a 234 nm. A separação cromatográfica foi obtida no tempo de 15 minutos, sendo linear na faixa de concentração de 0,5-200 Hg/mL. O método foi aplicado para análise simultânea de ezetimiba e sinvastatina em produtos farmacêuticos comerciais. Paralelamente, desenvolveu-se e validou-se método por cromatografia líquida combinada à espectrometria de massas (CL-EM/EM). Executaram-se as análises utilizando coluna Luna C18 (50 mm x 4,6 mm) e fase móvel composta de acetonitrila:água (85:15, V/V) na vazão de 0,4 mL/min. O espectrômetro de massas, equipado com fonte de electrospray positivo, foi empregado no modo de monitoramento de reação múltipla (MRM), monitorando as transições de 392,0>161,0 e 359,3>280,0, para a ezetimiba e etoricoxibe (padrão interno), respectivamente. A separação cromatográfica foi obtida em 2 minutos, sendo linear nas faixas de concentração de 0,25-20 ng/mL (ezetimiba) e 1-300 ng/mL (ezetimiba e seu metabólito). Os procedimentos foram validados, avaliando-se os parâmetros de especificidade, linearidade, precisão, exatidão, robustez, limite de detecção e quantificação, incluindo para o método bioanalítico os efeitos de matriz, recuperação e estudos de estabilidade, cujos resultados cumpriram os requisitos preconizados. O método proposto foi utilizado na análise de produtos farmacêuticos, demonstrando correlação significativa dos resultados (P>0,05). Além disso, o método de extração líquido-líquido desenvolvido e otimizado propiciou significativa percentagem de recuperação da ezetimiba, seu metabólito e do padrão interno nas amostras de plasma. Os procedimentos pesquisados podem ser aplicados para estudos de biodisponibilidade e para aprimorar o controle da qualidade de medicamentos.Universidade Federal de Santa MariaBRFarmáciaUFSMPrograma de Pós-Graduação em Ciências FarmacêuticasDalmora, Sergio Luizhttp://lattes.cnpq.br/4505166045049607Macedo, Rui Oliveirahttp://lattes.cnpq.br/8326594695097434Soares, Carlos Roberto Jorgehttp://lattes.cnpq.br/3805602544840562Oliveira, Paulo Renato de2007-10-232007-10-232007-03-30info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfOLIVEIRA, Paulo Renato de. Development and validation of methodologies for the evaluation of ezetimibe by liquid chromatography and mass spectrometry. 2007. 82 f. Dissertação (Mestrado em Farmacologia) - Universidade Federal de Santa Maria, Santa Maria, 2007.http://repositorio.ufsm.br/handle/1/6052porinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2022-10-17T14:50:41Zoai:repositorio.ufsm.br:1/6052Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2022-10-17T14:50:41Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Desenvolvimento e validação de metodologias para avaliação de ezetimiba por cromatografia líquida e espectrometria de massas Development and validation of methodologies for the evaluation of ezetimibe by liquid chromatography and mass spectrometry |
title |
Desenvolvimento e validação de metodologias para avaliação de ezetimiba por cromatografia líquida e espectrometria de massas |
spellingShingle |
Desenvolvimento e validação de metodologias para avaliação de ezetimiba por cromatografia líquida e espectrometria de massas Oliveira, Paulo Renato de Cromatografia líquida Espectrometria de massas Ezetimiba Validação Plasma humano Produtos farmacêuticos Liquid chromatography Mass spectrometry Ezetimibe Validation Human plasma Pharmaceutical products CNPQ::CIENCIAS DA SAUDE::FARMACIA |
title_short |
Desenvolvimento e validação de metodologias para avaliação de ezetimiba por cromatografia líquida e espectrometria de massas |
title_full |
Desenvolvimento e validação de metodologias para avaliação de ezetimiba por cromatografia líquida e espectrometria de massas |
title_fullStr |
Desenvolvimento e validação de metodologias para avaliação de ezetimiba por cromatografia líquida e espectrometria de massas |
title_full_unstemmed |
Desenvolvimento e validação de metodologias para avaliação de ezetimiba por cromatografia líquida e espectrometria de massas |
title_sort |
Desenvolvimento e validação de metodologias para avaliação de ezetimiba por cromatografia líquida e espectrometria de massas |
author |
Oliveira, Paulo Renato de |
author_facet |
Oliveira, Paulo Renato de |
author_role |
author |
dc.contributor.none.fl_str_mv |
Dalmora, Sergio Luiz http://lattes.cnpq.br/4505166045049607 Macedo, Rui Oliveira http://lattes.cnpq.br/8326594695097434 Soares, Carlos Roberto Jorge http://lattes.cnpq.br/3805602544840562 |
dc.contributor.author.fl_str_mv |
Oliveira, Paulo Renato de |
dc.subject.por.fl_str_mv |
Cromatografia líquida Espectrometria de massas Ezetimiba Validação Plasma humano Produtos farmacêuticos Liquid chromatography Mass spectrometry Ezetimibe Validation Human plasma Pharmaceutical products CNPQ::CIENCIAS DA SAUDE::FARMACIA |
topic |
Cromatografia líquida Espectrometria de massas Ezetimiba Validação Plasma humano Produtos farmacêuticos Liquid chromatography Mass spectrometry Ezetimibe Validation Human plasma Pharmaceutical products CNPQ::CIENCIAS DA SAUDE::FARMACIA |
description |
Ezetimibe selectively inhibits the intestinal absorption of dietary cholesterol and related plant sterols, from the 2-azetidinones group, and is used for the treatment of hypercholesterolemia and phytosterolemia. The methodologies for the evaluation of ezetimibe in pharmaceutical products and plasma were developed and validated in the present work. The reversed-phase liquid chromatography (RP-LC) analysis was carried out using a Synergi fusion C18 column (150 mm x 4.6 mm), maintained at 45 oC. The mobile phase consisted of potassium phosphate buffer 0.03 M, pH 4.5/acetonitrile (35:65, V/V), run at a flow rate of 0.6 mL/min with detection at 234 nm. The chromatographic separation was obtained within 15 min and it was linear in the concentration range of 0.5-200 Hg/mL. The method was sucessfuly applied for the simultaneous determination of ezetimibe and simvastatin in pharmaceutical products. The liquid chromatography-tandem mass spectrometry (LCMS/ MS) method was developed and validated using a Luna C18 column (50 mm x 3.0 mm) and the mobile phase consisted of acetonitrile:water (85:15, V/V), run at a flow rate of 0.4 mL/min. The mass spectrometer, equipped with electrospray positive source, was used in multiple reaction monitoring mode (MRM), monitoring the transitions of 392.0>161.0 and 359.3>280.0, for ezetimibe and etoricoxib (internal standard), respectively. The chromatographic separation was obtained within 2 min and it was linear in the concentration range of 0.25-20 ng/mL (ezetimibe) and 1-300 ng/mL (ezetimibe and its glucuronide matabolite). The procedures were validated evaluating parameters such as the specificity, linearity, precision, accuracy, robustness, limit of detection and limit of quantitation. Besides, for the bioanalytical method, the matrix effects, recovery and stability studies were also analyzed, giving results within the acceptable range. The proposed method was applied for the analysis of pharmaceutical products, showing significant correlation (P>0.05) of the results. Moreover, the liquid-liquid extraction method developed and optimized allowed high mean recoveries of ezetimibe and internal standard from the plasma samples. The procedures can be applied for the biovailability studies and for the quality control of pharmaceutical products. |
publishDate |
2007 |
dc.date.none.fl_str_mv |
2007-10-23 2007-10-23 2007-03-30 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
OLIVEIRA, Paulo Renato de. Development and validation of methodologies for the evaluation of ezetimibe by liquid chromatography and mass spectrometry. 2007. 82 f. Dissertação (Mestrado em Farmacologia) - Universidade Federal de Santa Maria, Santa Maria, 2007. http://repositorio.ufsm.br/handle/1/6052 |
identifier_str_mv |
OLIVEIRA, Paulo Renato de. Development and validation of methodologies for the evaluation of ezetimibe by liquid chromatography and mass spectrometry. 2007. 82 f. Dissertação (Mestrado em Farmacologia) - Universidade Federal de Santa Maria, Santa Maria, 2007. |
url |
http://repositorio.ufsm.br/handle/1/6052 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria BR Farmácia UFSM Programa de Pós-Graduação em Ciências Farmacêuticas |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria BR Farmácia UFSM Programa de Pós-Graduação em Ciências Farmacêuticas |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
_version_ |
1805922055357464576 |