Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD
Autor(a) principal: | |
---|---|
Data de Publicação: | 2013 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFRN |
Texto Completo: | https://repositorio.ufrn.br/jspui/handle/123456789/13477 |
Resumo: | Tuberculosis is a serious disease, but curable in practically 100% of new cases, since complied the principles of modern chemotherapy. Isoniazid (ISN), Rifampicin (RIF), Pyrazinamide (PYR) and Chloride Ethambutol (ETA) are considered first line drugs in the treatment of tuberculosis, by combining the highest level of efficiency with acceptable degree of toxicity. Concerning USP 33 - NF28 (2010) the chromatography analysis to 3 of 4 drugs (ISN, PYR and RIF) last in average 15 minutes and 10 minutes more to obtain the 4th drug (ETA) using a column and mobile phase mixture different, becoming its industrial application unfavorable. Thus, many studies have being carried out to minimize this problem. An alternative would use the UFLC, which is based with the same principles of HPLC, however it uses stationary phases with particles smaller than 2 μm. Therefore, this study goals to develop and validate new analytical methods to determine simultaneously the drugs by HPLC/DAD and UFLC/DAD. For this, a analytical screening was carried out, which verified that is necessary a gradient of mobile phase system A (acetate buffer:methanol 94:6 v/v) and B (acetate buffer:acetonitrile 55:45 v/v). Furthermore, to the development and optimization of the method in HPLC and UFLC, with achievement of the values of system suitability into the criteria limits required for both techniques, the validations have began. Standard solutions and tablets test solutions were prepared and injected into HPLC and UFLC, containing 0.008 mg/mL ISN, 0.043 mg/mL PYR, 0.030 mg.mL-1 ETA and 0.016 mg/mL RIF. The validation of analytical methods for HPLC and UFLC was carried out with the determination of specificity/selectivity, analytical curve, linearity, precision, limits of detection and quantification, accuracy and robustness. The methods were adequate for determination of 4 drugs separately without interfered with the others. Precise, due to the fact of the methods demonstrated since with the days variation, besides the repeatability, the values were into the level required by the regular agency. Linear (R> 0,99), once the methods were capable to demonstrate results directly proportional to the concentration of the analyte sample, within of specified range. Accurate, once the methods were capable to present values of variation coefficient and recovery percentage into the required limits (98 to 102%). The methods showed LOD and LOQ very low showing the high sensitivity of the methods for the four drugs. The robustness of the methods were evaluate, facing the temperature and flow changes, where they showed robustness just with the preview conditions established of temperature and flow, abrupt changes may influence with the results of methods |
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Paiva, Marcelo Vitor dehttp://lattes.cnpq.br/6323471692543395http://lattes.cnpq.br/9657118649043311Gomes, Ana Paula Barretohttp://lattes.cnpq.br/1689823596741892Pianetti, Gerson Antoniohttp://lattes.cnpq.br/5212837629822557Aragão, Cícero Flávio Soares2014-12-17T14:16:30Z2013-05-092014-12-17T14:16:30Z2013-03-21PAIVA, Marcelo Vitor de. Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD. 2013. 105 f. Dissertação (Mestrado em Bioanálises e Medicamentos) - Universidade Federal do Rio Grande do Norte, Natal, 2013.https://repositorio.ufrn.br/jspui/handle/123456789/13477Tuberculosis is a serious disease, but curable in practically 100% of new cases, since complied the principles of modern chemotherapy. Isoniazid (ISN), Rifampicin (RIF), Pyrazinamide (PYR) and Chloride Ethambutol (ETA) are considered first line drugs in the treatment of tuberculosis, by combining the highest level of efficiency with acceptable degree of toxicity. Concerning USP 33 - NF28 (2010) the chromatography analysis to 3 of 4 drugs (ISN, PYR and RIF) last in average 15 minutes and 10 minutes more to obtain the 4th drug (ETA) using a column and mobile phase mixture different, becoming its industrial application unfavorable. Thus, many studies have being carried out to minimize this problem. An alternative would use the UFLC, which is based with the same principles of HPLC, however it uses stationary phases with particles smaller than 2 μm. Therefore, this study goals to develop and validate new analytical methods to determine simultaneously the drugs by HPLC/DAD and UFLC/DAD. For this, a analytical screening was carried out, which verified that is necessary a gradient of mobile phase system A (acetate buffer:methanol 94:6 v/v) and B (acetate buffer:acetonitrile 55:45 v/v). Furthermore, to the development and optimization of the method in HPLC and UFLC, with achievement of the values of system suitability into the criteria limits required for both techniques, the validations have began. Standard solutions and tablets test solutions were prepared and injected into HPLC and UFLC, containing 0.008 mg/mL ISN, 0.043 mg/mL PYR, 0.030 mg.mL-1 ETA and 0.016 mg/mL RIF. The validation of analytical methods for HPLC and UFLC was carried out with the determination of specificity/selectivity, analytical curve, linearity, precision, limits of detection and quantification, accuracy and robustness. The methods were adequate for determination of 4 drugs separately without interfered with the others. Precise, due to the fact of the methods demonstrated since with the days variation, besides the repeatability, the values were into the level required by the regular agency. Linear (R> 0,99), once the methods were capable to demonstrate results directly proportional to the concentration of the analyte sample, within of specified range. Accurate, once the methods were capable to present values of variation coefficient and recovery percentage into the required limits (98 to 102%). The methods showed LOD and LOQ very low showing the high sensitivity of the methods for the four drugs. The robustness of the methods were evaluate, facing the temperature and flow changes, where they showed robustness just with the preview conditions established of temperature and flow, abrupt changes may influence with the results of methodsA tuberculose é uma doença grave, porém curável em praticamente 100% dos casos novos, desde que obedecidos os princípios da moderna quimioterapia. São considerados fármacos de 1ª linha no tratamento à tuberculose: isoniazida, pirazinamida, etambutol e rifampicina. De acordo com USP 33 - NF28 (2010) as análises cromatográficas para 3 dos 4 fármacos (isoniazida, pirazinamida e rifampicina) duram em média 15 minutos e mais 10 minutos para a obtenção do 4° fármaco (etambutol) utilizando outra coluna, com outra mistura de fase móvel, tornando esta aplicação na prática industrial desfavorável. Uma das alternativas é utilizar o CLUE, o qual baseia-se nos mesmos princípios da CLAE, porém utiliza fases estacionárias com partículas menores que 2 μm. Dessa forma pretende-se com o presente estudo desenvolver e validar novos métodos analíticos para determinação simultânea de tuberculostáticos por CLAE/DAD e CLUE/DAD. Para isto, foi realizado um screening analítico, o qual verificou que é necessário um gradiente de sistema de fase móvel A (tampão acetato:metanol 94:6 v/v) e B (tampão acetato:acetonitrila 55:45 v/v). Posteriormente, ao desenvolvimento e otimização do método em CLAE e CLUE com a obtenção dos valores de adequabilidade do sistema dentro dos limites de aceitações vigente para ambos as técnicas, as validações deram-se início. Soluções padrões e soluções testes dos comprimidos foram preparadas e injetadas no CLAE e CLUE, contendo isoniazida, pirazinamida, etambutol e rifampicina nas concentrações de 0,008, 0,043, 0,030 e 0,016 mg.mL-1, respectivamente. A validação dos métodos analíticos foram realizadas para: especificidade / seletividade, intervalos da curva analítica, linearidade, limite de detecção, limite de quantificação, exatidão, precisão (repetibilidade, precisão intermediária) e robustez. Os métodos foram adequados para determinação dos 4 fármacos separadamente sem interferência nos demais. Precisos, devido ao fato de que os métodos demonstraram que mesmo com variação de dias, além da repetibilidade, os valores ficaram dentro da faixa preconizada na legislação vigente. Lineares (R > 0,99), ou seja, os métodos foram capazes de demonstrar que os resultados obtidos eram diretamente proporcionais à concentração do analito na amostra, dentro de um intervalo especificado. Exatos, uma vez que os métodos foram capazes de apresentar valores de coeficiente de variação e porcentagem de recuperação dentro dos limites exigidos (98 a 102%). Os métodos mostraram LD e LQ com níveis baixos demonstrando que os métodos possuem elevada sensibilidade aos quarto fármacos. A robustez foi avaliada frente às alterações de temperatura e fluxo, onde os métodos demonstraram-se robustos apenas nas condições previamente estabelecidas de temperatura e fluxo, alterações bruscas podem acarretar influência nos resultados dos métodosapplication/pdfporUniversidade Federal do Rio Grande do NortePrograma de Pós-Graduação em Ciências FarmacêuticasUFRNBRBioanálises e MedicamentosIsoniazida. Pirazinamida. Etambutol. Rifampicina. Cromatografia Líquida de Alta Eficiência. Cromatografia Líquida de Ultra EficiênciaIsoniazid. Pyrazinamide. Ethambutol. Rifampicin. High Performance Liquid Chromatography. 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dc.title.por.fl_str_mv |
Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD |
title |
Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD |
spellingShingle |
Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD Paiva, Marcelo Vitor de Isoniazida. Pirazinamida. Etambutol. Rifampicina. Cromatografia Líquida de Alta Eficiência. Cromatografia Líquida de Ultra Eficiência Isoniazid. Pyrazinamide. Ethambutol. Rifampicin. High Performance Liquid Chromatography. Ultra Fast Liquid Chromatography CNPQ::OUTROS |
title_short |
Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD |
title_full |
Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD |
title_fullStr |
Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD |
title_full_unstemmed |
Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD |
title_sort |
Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD |
author |
Paiva, Marcelo Vitor de |
author_facet |
Paiva, Marcelo Vitor de |
author_role |
author |
dc.contributor.authorID.por.fl_str_mv |
|
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/6323471692543395 |
dc.contributor.advisorID.por.fl_str_mv |
|
dc.contributor.advisorLattes.por.fl_str_mv |
http://lattes.cnpq.br/9657118649043311 |
dc.contributor.referees1.pt_BR.fl_str_mv |
Gomes, Ana Paula Barreto |
dc.contributor.referees1ID.por.fl_str_mv |
|
dc.contributor.referees1Lattes.por.fl_str_mv |
http://lattes.cnpq.br/1689823596741892 |
dc.contributor.referees2.pt_BR.fl_str_mv |
Pianetti, Gerson Antonio |
dc.contributor.referees2ID.por.fl_str_mv |
|
dc.contributor.referees2Lattes.por.fl_str_mv |
http://lattes.cnpq.br/5212837629822557 |
dc.contributor.author.fl_str_mv |
Paiva, Marcelo Vitor de |
dc.contributor.advisor1.fl_str_mv |
Aragão, Cícero Flávio Soares |
contributor_str_mv |
Aragão, Cícero Flávio Soares |
dc.subject.por.fl_str_mv |
Isoniazida. Pirazinamida. Etambutol. Rifampicina. Cromatografia Líquida de Alta Eficiência. Cromatografia Líquida de Ultra Eficiência |
topic |
Isoniazida. Pirazinamida. Etambutol. Rifampicina. Cromatografia Líquida de Alta Eficiência. Cromatografia Líquida de Ultra Eficiência Isoniazid. Pyrazinamide. Ethambutol. Rifampicin. High Performance Liquid Chromatography. Ultra Fast Liquid Chromatography CNPQ::OUTROS |
dc.subject.eng.fl_str_mv |
Isoniazid. Pyrazinamide. Ethambutol. Rifampicin. High Performance Liquid Chromatography. Ultra Fast Liquid Chromatography |
dc.subject.cnpq.fl_str_mv |
CNPQ::OUTROS |
description |
Tuberculosis is a serious disease, but curable in practically 100% of new cases, since complied the principles of modern chemotherapy. Isoniazid (ISN), Rifampicin (RIF), Pyrazinamide (PYR) and Chloride Ethambutol (ETA) are considered first line drugs in the treatment of tuberculosis, by combining the highest level of efficiency with acceptable degree of toxicity. Concerning USP 33 - NF28 (2010) the chromatography analysis to 3 of 4 drugs (ISN, PYR and RIF) last in average 15 minutes and 10 minutes more to obtain the 4th drug (ETA) using a column and mobile phase mixture different, becoming its industrial application unfavorable. Thus, many studies have being carried out to minimize this problem. An alternative would use the UFLC, which is based with the same principles of HPLC, however it uses stationary phases with particles smaller than 2 μm. Therefore, this study goals to develop and validate new analytical methods to determine simultaneously the drugs by HPLC/DAD and UFLC/DAD. For this, a analytical screening was carried out, which verified that is necessary a gradient of mobile phase system A (acetate buffer:methanol 94:6 v/v) and B (acetate buffer:acetonitrile 55:45 v/v). Furthermore, to the development and optimization of the method in HPLC and UFLC, with achievement of the values of system suitability into the criteria limits required for both techniques, the validations have began. Standard solutions and tablets test solutions were prepared and injected into HPLC and UFLC, containing 0.008 mg/mL ISN, 0.043 mg/mL PYR, 0.030 mg.mL-1 ETA and 0.016 mg/mL RIF. The validation of analytical methods for HPLC and UFLC was carried out with the determination of specificity/selectivity, analytical curve, linearity, precision, limits of detection and quantification, accuracy and robustness. The methods were adequate for determination of 4 drugs separately without interfered with the others. Precise, due to the fact of the methods demonstrated since with the days variation, besides the repeatability, the values were into the level required by the regular agency. Linear (R> 0,99), once the methods were capable to demonstrate results directly proportional to the concentration of the analyte sample, within of specified range. Accurate, once the methods were capable to present values of variation coefficient and recovery percentage into the required limits (98 to 102%). The methods showed LOD and LOQ very low showing the high sensitivity of the methods for the four drugs. The robustness of the methods were evaluate, facing the temperature and flow changes, where they showed robustness just with the preview conditions established of temperature and flow, abrupt changes may influence with the results of methods |
publishDate |
2013 |
dc.date.available.fl_str_mv |
2013-05-09 2014-12-17T14:16:30Z |
dc.date.issued.fl_str_mv |
2013-03-21 |
dc.date.accessioned.fl_str_mv |
2014-12-17T14:16:30Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
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PAIVA, Marcelo Vitor de. Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD. 2013. 105 f. Dissertação (Mestrado em Bioanálises e Medicamentos) - Universidade Federal do Rio Grande do Norte, Natal, 2013. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufrn.br/jspui/handle/123456789/13477 |
identifier_str_mv |
PAIVA, Marcelo Vitor de. Otimização e validação de métodos analíticos para a determinação simultânea de tuberculostáticos (4-FDC) por CLAE/DAD e CLUE/ DAD. 2013. 105 f. Dissertação (Mestrado em Bioanálises e Medicamentos) - Universidade Federal do Rio Grande do Norte, Natal, 2013. |
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https://repositorio.ufrn.br/jspui/handle/123456789/13477 |
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Universidade Federal do Rio Grande do Norte |
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Programa de Pós-Graduação em Ciências Farmacêuticas |
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UFRN |
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BR |
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Bioanálises e Medicamentos |
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Universidade Federal do Rio Grande do Norte |
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