Metabolismo in vitro do albendazol em escala miligrama e purificação por cromatografia líquida de alta eficiência semipreparativa
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/6572 |
Resumo: | This work presents the in vitro metabolism of albendazole (ABZ) conducted with microsomal fractions obtained from Wistar rats and performed in analytical and semipreparative scale. Subsequent separation and purification of the major metabolites was accomplished using High Performance Chiral Liquid Chromatography in normal elution mode. The chromatographic separation of ABZ, albendazol sulphoxide (ABZ-SO), albendazole 2-aminesulphone (ABZ-NH2-SO2) and albendazole sulphone (ABZ-SO2) was achieved on a chromatographic chiral column containing amylose tris(3,5-dimethylphenylcarbamate) (15.0 x 0.46 cm i.d.) or (20.0 x 0.70 cm i.d.), hexane:ethanol (55:45) as mobile phase and = 290 nm. Different parameters in the in vitro biotransformation were evaluated in a univariate mode and the best conditions selected as follows: incubation time (6 hours), concentration of ABZ (1.5 mg/mL), concentration of proteins (20 mg/ml) and concentration of reduced nicotinamide adenine dinucleotide phosphate dehydrogenase (NADPH; 20 mg/ml). For the NADPH regenerating system was used nicotinamide adenine dinucleotide phosphate (NADP) 100 mmol/L, glucose-6-phosphate (G6P) 100 mmol/L and glucose-6-phosphate dehydrogenase (G6PD) 100 U/mL. For the microsomal fractions both the calcium aggregation or ultracentrifugation methods were appropriate for microsome isolation. Ultracentrifugation provided the highest protein concentration for the extracted samples (35.2 mg/mL). The sample preparation was conducted by solid phase extraction and the recoveries were within 89 to 109%. The in vitro metabolism in milligram scale allowed the production of 1.0 mg of the (+)- ABZ-SO and 8.0 mg of the (-)-ABZ-SO. The ABZ-NH2-SO2 and ABZ-SO2 were not observed under the biotransformation conditions tested. The in vitro metabolism in milligram scale can be used for the production and purification of metabolites for further pharmacokinetics studies. |
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Batistão, Marina BeghiniOliveira, Regina Vincenzihttp://lattes.cnpq.br/6609377714413073http://lattes.cnpq.br/842589388543027434a2f4c9-5dba-4c0b-af75-c9d9a4e9ea4a2016-06-02T20:36:48Z2013-12-172016-06-02T20:36:48Z2012-06-06BATISTÃO, Marina Beghini. In vitro metabolism of albendazole in milligram scale and purification by semipreparative high perfomance liquid chromatography. 2012. 122 f. Dissertação (Mestrado em Ciências Exatas e da Terra) - Universidade Federal de São Carlos, São Carlos, 2012.https://repositorio.ufscar.br/handle/ufscar/6572This work presents the in vitro metabolism of albendazole (ABZ) conducted with microsomal fractions obtained from Wistar rats and performed in analytical and semipreparative scale. Subsequent separation and purification of the major metabolites was accomplished using High Performance Chiral Liquid Chromatography in normal elution mode. The chromatographic separation of ABZ, albendazol sulphoxide (ABZ-SO), albendazole 2-aminesulphone (ABZ-NH2-SO2) and albendazole sulphone (ABZ-SO2) was achieved on a chromatographic chiral column containing amylose tris(3,5-dimethylphenylcarbamate) (15.0 x 0.46 cm i.d.) or (20.0 x 0.70 cm i.d.), hexane:ethanol (55:45) as mobile phase and = 290 nm. Different parameters in the in vitro biotransformation were evaluated in a univariate mode and the best conditions selected as follows: incubation time (6 hours), concentration of ABZ (1.5 mg/mL), concentration of proteins (20 mg/ml) and concentration of reduced nicotinamide adenine dinucleotide phosphate dehydrogenase (NADPH; 20 mg/ml). For the NADPH regenerating system was used nicotinamide adenine dinucleotide phosphate (NADP) 100 mmol/L, glucose-6-phosphate (G6P) 100 mmol/L and glucose-6-phosphate dehydrogenase (G6PD) 100 U/mL. For the microsomal fractions both the calcium aggregation or ultracentrifugation methods were appropriate for microsome isolation. Ultracentrifugation provided the highest protein concentration for the extracted samples (35.2 mg/mL). The sample preparation was conducted by solid phase extraction and the recoveries were within 89 to 109%. The in vitro metabolism in milligram scale allowed the production of 1.0 mg of the (+)- ABZ-SO and 8.0 mg of the (-)-ABZ-SO. The ABZ-NH2-SO2 and ABZ-SO2 were not observed under the biotransformation conditions tested. The in vitro metabolism in milligram scale can be used for the production and purification of metabolites for further pharmacokinetics studies.Este trabalho apresenta o metabolismo in vitro do albendazol (ABZ), realizado em frações microssomais de fígados de ratos Wistar, em escala analítica e semipreparativa. Posteriormente, a separação e isolamento dos metabólitos majoritários foi realizada com emprego de Cromatografia Liquida Quiral de Alta Eficiência no modo normal de eluição. A separação cromatográfica do ABZ, albendazol sulfóxido (ABZ-SO), albendazol 2-aminosulfona (ABZ-NH2-SO2) e albendazol sulfona (ABZ-SO2) foi obtida utilizando uma coluna cromatográfica quiral tris(3,5-dimetilfenilcarbamato) de amilose em sílica APS-Nucleosil (15,0 x 0,46 cm d.i.) ou (20,0 x 0,70 cm d.i.), fase móvel composta por hexano:etanol (55:45 v/v) e =290 nm. Diferentes parâmetros do processo de biotransformação in vitro foram avaliados, no modo univariado, e as melhores condições selecionadas, sendo estas: tempo de incubação (6 horas), concentração do ABZ (1,5 mg/mL), concentração proteica (20 mg/mL) e concentração de ß-nicotinamida-adenina-dinucleotídeoreduzida (NADPH; 20 mg/mL). Para o sistema regenerador de NADPH, utilizou-se ßnicotinamida- adenina-dinucleotídeo fosfato (NADP) 100 mmol/L, glicose-6-fosfato (G6P) 100 mmol/L e glicose-6-fosfato desidrogenase (G6PD) 100 U/mL. Para a obtenção das frações microssomais, tanto o método de precipitação com cloreto de cálcio quanto o de ultracentrifugação, foram apropriados para o isolamento dos microssomas. A ultracentrifugação propiciou uma maior concentração protéica para as amostras extraídas (35,2 mg/mL). O preparo das amostras foi realizado por extração em fase sólida, com valores de recuperação de 89 a 109%. O metabolismo in vitro em escala miligrama propiciou a obtenção de 1,0 mg do (+)-ABZ-SO e 8,0 mg do (-)-ABZ-SO. O ABZ-NH2-SO2 e ABZ-SO2 não foram observados nas condições de biotransformação avaliadas. O metabolismo in vitro em escala miligrama pode ser utilizado para a produção e isolamento de metabólitos para posterior estudos farmacocinéticos.Financiadora de Estudos e Projetosapplication/pdfporUniversidade Federal de São CarlosPrograma de Pós-Graduação em Química - PPGQUFSCarBRQuímica analíticaMetabolismo in vitroBenzimidazóisCromatografia quiralCIENCIAS EXATAS E DA TERRA::QUIMICAMetabolismo in vitro do albendazol em escala miligrama e purificação por cromatografia líquida de alta eficiência semipreparativaIn vitro metabolism of albendazole in milligram scale and purification by semipreparative high perfomance liquid chromatographyinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis-1-1c952ec5c-f2b5-49c6-8523-21d001220087info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINAL5628.pdfapplication/pdf1395038https://repositorio.ufscar.br/bitstream/ufscar/6572/1/5628.pdf50d9a83b9a58334f0e8e2735f43b4abdMD51TEXT5628.pdf.txt5628.pdf.txtExtracted texttext/plain0https://repositorio.ufscar.br/bitstream/ufscar/6572/4/5628.pdf.txtd41d8cd98f00b204e9800998ecf8427eMD54THUMBNAIL5628.pdf.jpg5628.pdf.jpgIM Thumbnailimage/jpeg8925https://repositorio.ufscar.br/bitstream/ufscar/6572/5/5628.pdf.jpgc3a1e281359237f4f782bc530ec1579cMD55ufscar/65722023-09-18 18:30:39.677oai:repositorio.ufscar.br:ufscar/6572Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:30:39Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Metabolismo in vitro do albendazol em escala miligrama e purificação por cromatografia líquida de alta eficiência semipreparativa |
dc.title.alternative.eng.fl_str_mv |
In vitro metabolism of albendazole in milligram scale and purification by semipreparative high perfomance liquid chromatography |
title |
Metabolismo in vitro do albendazol em escala miligrama e purificação por cromatografia líquida de alta eficiência semipreparativa |
spellingShingle |
Metabolismo in vitro do albendazol em escala miligrama e purificação por cromatografia líquida de alta eficiência semipreparativa Batistão, Marina Beghini Química analítica Metabolismo in vitro Benzimidazóis Cromatografia quiral CIENCIAS EXATAS E DA TERRA::QUIMICA |
title_short |
Metabolismo in vitro do albendazol em escala miligrama e purificação por cromatografia líquida de alta eficiência semipreparativa |
title_full |
Metabolismo in vitro do albendazol em escala miligrama e purificação por cromatografia líquida de alta eficiência semipreparativa |
title_fullStr |
Metabolismo in vitro do albendazol em escala miligrama e purificação por cromatografia líquida de alta eficiência semipreparativa |
title_full_unstemmed |
Metabolismo in vitro do albendazol em escala miligrama e purificação por cromatografia líquida de alta eficiência semipreparativa |
title_sort |
Metabolismo in vitro do albendazol em escala miligrama e purificação por cromatografia líquida de alta eficiência semipreparativa |
author |
Batistão, Marina Beghini |
author_facet |
Batistão, Marina Beghini |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/8425893885430274 |
dc.contributor.author.fl_str_mv |
Batistão, Marina Beghini |
dc.contributor.advisor1.fl_str_mv |
Oliveira, Regina Vincenzi |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/6609377714413073 |
dc.contributor.authorID.fl_str_mv |
34a2f4c9-5dba-4c0b-af75-c9d9a4e9ea4a |
contributor_str_mv |
Oliveira, Regina Vincenzi |
dc.subject.por.fl_str_mv |
Química analítica Metabolismo in vitro Benzimidazóis Cromatografia quiral |
topic |
Química analítica Metabolismo in vitro Benzimidazóis Cromatografia quiral CIENCIAS EXATAS E DA TERRA::QUIMICA |
dc.subject.cnpq.fl_str_mv |
CIENCIAS EXATAS E DA TERRA::QUIMICA |
description |
This work presents the in vitro metabolism of albendazole (ABZ) conducted with microsomal fractions obtained from Wistar rats and performed in analytical and semipreparative scale. Subsequent separation and purification of the major metabolites was accomplished using High Performance Chiral Liquid Chromatography in normal elution mode. The chromatographic separation of ABZ, albendazol sulphoxide (ABZ-SO), albendazole 2-aminesulphone (ABZ-NH2-SO2) and albendazole sulphone (ABZ-SO2) was achieved on a chromatographic chiral column containing amylose tris(3,5-dimethylphenylcarbamate) (15.0 x 0.46 cm i.d.) or (20.0 x 0.70 cm i.d.), hexane:ethanol (55:45) as mobile phase and = 290 nm. Different parameters in the in vitro biotransformation were evaluated in a univariate mode and the best conditions selected as follows: incubation time (6 hours), concentration of ABZ (1.5 mg/mL), concentration of proteins (20 mg/ml) and concentration of reduced nicotinamide adenine dinucleotide phosphate dehydrogenase (NADPH; 20 mg/ml). For the NADPH regenerating system was used nicotinamide adenine dinucleotide phosphate (NADP) 100 mmol/L, glucose-6-phosphate (G6P) 100 mmol/L and glucose-6-phosphate dehydrogenase (G6PD) 100 U/mL. For the microsomal fractions both the calcium aggregation or ultracentrifugation methods were appropriate for microsome isolation. Ultracentrifugation provided the highest protein concentration for the extracted samples (35.2 mg/mL). The sample preparation was conducted by solid phase extraction and the recoveries were within 89 to 109%. The in vitro metabolism in milligram scale allowed the production of 1.0 mg of the (+)- ABZ-SO and 8.0 mg of the (-)-ABZ-SO. The ABZ-NH2-SO2 and ABZ-SO2 were not observed under the biotransformation conditions tested. The in vitro metabolism in milligram scale can be used for the production and purification of metabolites for further pharmacokinetics studies. |
publishDate |
2012 |
dc.date.issued.fl_str_mv |
2012-06-06 |
dc.date.available.fl_str_mv |
2013-12-17 2016-06-02T20:36:48Z |
dc.date.accessioned.fl_str_mv |
2016-06-02T20:36:48Z |
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.citation.fl_str_mv |
BATISTÃO, Marina Beghini. In vitro metabolism of albendazole in milligram scale and purification by semipreparative high perfomance liquid chromatography. 2012. 122 f. Dissertação (Mestrado em Ciências Exatas e da Terra) - Universidade Federal de São Carlos, São Carlos, 2012. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/6572 |
identifier_str_mv |
BATISTÃO, Marina Beghini. In vitro metabolism of albendazole in milligram scale and purification by semipreparative high perfomance liquid chromatography. 2012. 122 f. Dissertação (Mestrado em Ciências Exatas e da Terra) - Universidade Federal de São Carlos, São Carlos, 2012. |
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https://repositorio.ufscar.br/handle/ufscar/6572 |
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Universidade Federal de São Carlos |
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Programa de Pós-Graduação em Química - PPGQ |
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UFSCar |
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BR |
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Universidade Federal de São Carlos |
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