Determinação de benzodiazepínicos em plasma por cromatografia líquida com detector ultravioleta-visível empregando microextração líquido-líquido dispersiva
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
dARK ID: | ark:/26339/0013000009vd5 |
Texto Completo: | http://repositorio.ufsm.br/handle/1/21163 |
Resumo: | Drugs has been the main agents involved in intoxications in Brazil. According to the latest report of SINITOX, these agents summed up 20367 cases representing 26,79% of all agents involved. Among them, 9983 cases were suicide attempts. Psychotropic drugs of the benzodiazepine class were the agents with bigger representativity in this scenario. Among them, Clonazepam (Rivotril®) was involved in 1506 cases (1223 under intentional circumstance). Benzodiazepines possess pharmacological characteristics convenient for suicide attempts and as drug facilitated crimes. As these characteristics are not exclusive of this drug class, it is necessary a development of an analytical method capable of detect and quantify these drugs with adequate specificity and sensitivity to help in the doctor behavior and other health professionals in accidental or intentional intoxications. In this context, blood is considered a complex matrix that allows to determine the concentration of this xenobiotic which is still active in the organism, allowing to correlate its levels with the biological effects, besides being a sample of choice for toxicological analysis. However, extractive procedures are necessary as previous step to promote biological matrix clean up, avoiding equipment damage and to other inputs as concentrate the analytes of interest allowing the chromatographic system detection. Among the extractive techniques, the dispersive liquid-liquid microextraction (DLLME) stands out for being a quick, simple, low cost process adaptable in any laboratory and it was little explored for the purpose of toxicological analyses. The main objective of this research was the development for an analytical method for the benzodiazepine analysis in plasma aiming to improve the medical behavior in accidental or intentional intoxications from the Hospital Universitário de Santa Maria located in the Universidade Federal de Santa Maria (HUSMUFSM). The results revealed that the extractive procedure got optimized this way, from 500 μL of plasma, previously alkalized to pH 10, are added 0,013 g of sodium chloride, then 400 μL of chloroform and 700 μL of acetonitrile are injected simultaneously, the content is homogenized in vortex for 10 seconds, then it is placed under ultrasonic bath por 1 minute, then the tube is centrifuged by 10000 rpm/10 minutes, the organic phase is collected, dried and resurrected with 30 μL of mobile phase (water pH 9/methanol/acetonitrile, 63:19:18) and 20 μL in the HPLCDAD. In the analytical validation, according to UNODC and SWGTOX guides, it wasn’t possible to validate all the parameters, it is possible to apply this method only for screening. |
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Determinação de benzodiazepínicos em plasma por cromatografia líquida com detector ultravioleta-visível empregando microextração líquido-líquido dispersivaDetermination of benzodiapines in plasma by high performance liquid chromatography with ultraviolet-visible detector applying dispersive liquid-liquid microextractionMicroextração líquido-líquido dispersivaBenzodiazepínicosPlasmaCromatografia líquidaIntoxicaçõesDispersive liquid-liquid microextractionLiquid chromatographyIntoxicationsCNPQ::CIENCIAS DA SAUDE::FARMACIADrugs has been the main agents involved in intoxications in Brazil. According to the latest report of SINITOX, these agents summed up 20367 cases representing 26,79% of all agents involved. Among them, 9983 cases were suicide attempts. Psychotropic drugs of the benzodiazepine class were the agents with bigger representativity in this scenario. Among them, Clonazepam (Rivotril®) was involved in 1506 cases (1223 under intentional circumstance). Benzodiazepines possess pharmacological characteristics convenient for suicide attempts and as drug facilitated crimes. As these characteristics are not exclusive of this drug class, it is necessary a development of an analytical method capable of detect and quantify these drugs with adequate specificity and sensitivity to help in the doctor behavior and other health professionals in accidental or intentional intoxications. In this context, blood is considered a complex matrix that allows to determine the concentration of this xenobiotic which is still active in the organism, allowing to correlate its levels with the biological effects, besides being a sample of choice for toxicological analysis. However, extractive procedures are necessary as previous step to promote biological matrix clean up, avoiding equipment damage and to other inputs as concentrate the analytes of interest allowing the chromatographic system detection. Among the extractive techniques, the dispersive liquid-liquid microextraction (DLLME) stands out for being a quick, simple, low cost process adaptable in any laboratory and it was little explored for the purpose of toxicological analyses. The main objective of this research was the development for an analytical method for the benzodiazepine analysis in plasma aiming to improve the medical behavior in accidental or intentional intoxications from the Hospital Universitário de Santa Maria located in the Universidade Federal de Santa Maria (HUSMUFSM). The results revealed that the extractive procedure got optimized this way, from 500 μL of plasma, previously alkalized to pH 10, are added 0,013 g of sodium chloride, then 400 μL of chloroform and 700 μL of acetonitrile are injected simultaneously, the content is homogenized in vortex for 10 seconds, then it is placed under ultrasonic bath por 1 minute, then the tube is centrifuged by 10000 rpm/10 minutes, the organic phase is collected, dried and resurrected with 30 μL of mobile phase (water pH 9/methanol/acetonitrile, 63:19:18) and 20 μL in the HPLCDAD. In the analytical validation, according to UNODC and SWGTOX guides, it wasn’t possible to validate all the parameters, it is possible to apply this method only for screening.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESOs medicamentos tem sido os principais agentes envolvidos em intoxicações no país. Segundo o último relatório do Sistema Nacional de Informações Toxico-Farmacológicas, estes agentes totalizaram 20367 casos de intoxicação representando 26,79% de todos os agentes envolvidos. Dentre este total, 9983 casos são de tentativa de suicídio. Os medicamentos psicotrópicos da classe dos benzodiazepínicos foram os agentes com maior representatividade neste cenário. Dentre os representantes, o Clonazepam (Rivotril®) se destacou em 1506 casos (1223 em circunstância intencional). Benzodiazepínicos possuem características farmacológicas as quais estão relacionadas à tentativas de suicídio e como facilitadores de crimes. Como estas características não são exclusivas desta classe de medicamentos, é necessário o desenvolvimento de um método analítico que possa detectar e quantificar estes fármacos com especificidade e sensibilidade suficientes para auxiliar na conduta profissional de médicos e outros profissionais da saúde em casos de intoxicação acidental ou intencional. Neste contexto, o sangue é considerado uma matriz complexa que permite avaliar a concentração deste xenobiótico que ainda está ativo no organismo, podendo correlacionar seus níveis com efeitos biológicos, além de ser uma amostra de escolha para fins toxicológicos. Entretanto, procedimentos extrativos são necessários como etapa anterior a injeção cromatográfica para promover o clean-up da matriz biológica, evitando danos ao equipamento e a outros insumos assim como concentrar os analitos de interesse para permitir a detecção pelo sistema cromatográfico. Entre as técnicas extrativas, a microextração líquido-líquido dispersiva (DLLME) destaca-se por se tratar de um processo rápido, simples, baixo custo e aplicável a qualquer laboratório e que foi pouco explorada para fins de análises toxicológicas. O objetivo geral desta pesquisa foi o desenvolvimento de um método analítico para análise de benzodiazepínicos em plasma visando auxiliar na conduta médica em casos de intoxicação acidental ou intencional oriundos do Hospital Universitário de Santa Maria localizado na Universidade Federal de Santa Maria (HUSM-UFSM). Os resultados mostraram que o procedimento extrativo ficou otimizado da seguinte forma, a partir de 500 μL de plasma, previamente alcalinizados a pH 10, são adicionados 0,013 g de cloreto de sódio, depois são injetados simultaneamente 400 μL de clorofórmio e 700 μL de acetonitrila, o conteúdo é homogeneizado em vórtex por 10 segundos e depois colocado banho ultrassônico por 1 minuto, depois centrifuga-se à 10000 rpm/10 minutos, a fase orgânica é coletada, evaporada e ressuspendida em 30 μL de fase móvel (água pH 9/metanol/acetonitrila, 63:19:18) e são injetados 20 μL no HPLC-DAD. Na validação analítica, segundo os guias da UNODC e SWGTOX, não foi possível validar todos os parâmetros, sendo possível a aplicação deste método apenas para triagem.Universidade Federal de Santa MariaBrasilAnálises Clínicas e ToxicológicasUFSMPrograma de Pós-Graduação em Ciências FarmacêuticasCentro de Ciências da SaúdeBairros, André Valle dehttp://lattes.cnpq.br/9350662167425345Oliveira, Sarah Carobini Werner De Souza Eller Franco dePrestes, Osmar DamianSaldanha, Geovane de Almeida2021-06-21T17:03:40Z2021-06-21T17:03:40Z2020-03-27info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/21163ark:/26339/0013000009vd5porAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2021-08-03T20:29:35Zoai:repositorio.ufsm.br:1/21163Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2021-08-03T20:29:35Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Determinação de benzodiazepínicos em plasma por cromatografia líquida com detector ultravioleta-visível empregando microextração líquido-líquido dispersiva Determination of benzodiapines in plasma by high performance liquid chromatography with ultraviolet-visible detector applying dispersive liquid-liquid microextraction |
title |
Determinação de benzodiazepínicos em plasma por cromatografia líquida com detector ultravioleta-visível empregando microextração líquido-líquido dispersiva |
spellingShingle |
Determinação de benzodiazepínicos em plasma por cromatografia líquida com detector ultravioleta-visível empregando microextração líquido-líquido dispersiva Saldanha, Geovane de Almeida Microextração líquido-líquido dispersiva Benzodiazepínicos Plasma Cromatografia líquida Intoxicações Dispersive liquid-liquid microextraction Liquid chromatography Intoxications CNPQ::CIENCIAS DA SAUDE::FARMACIA |
title_short |
Determinação de benzodiazepínicos em plasma por cromatografia líquida com detector ultravioleta-visível empregando microextração líquido-líquido dispersiva |
title_full |
Determinação de benzodiazepínicos em plasma por cromatografia líquida com detector ultravioleta-visível empregando microextração líquido-líquido dispersiva |
title_fullStr |
Determinação de benzodiazepínicos em plasma por cromatografia líquida com detector ultravioleta-visível empregando microextração líquido-líquido dispersiva |
title_full_unstemmed |
Determinação de benzodiazepínicos em plasma por cromatografia líquida com detector ultravioleta-visível empregando microextração líquido-líquido dispersiva |
title_sort |
Determinação de benzodiazepínicos em plasma por cromatografia líquida com detector ultravioleta-visível empregando microextração líquido-líquido dispersiva |
author |
Saldanha, Geovane de Almeida |
author_facet |
Saldanha, Geovane de Almeida |
author_role |
author |
dc.contributor.none.fl_str_mv |
Bairros, André Valle de http://lattes.cnpq.br/9350662167425345 Oliveira, Sarah Carobini Werner De Souza Eller Franco de Prestes, Osmar Damian |
dc.contributor.author.fl_str_mv |
Saldanha, Geovane de Almeida |
dc.subject.por.fl_str_mv |
Microextração líquido-líquido dispersiva Benzodiazepínicos Plasma Cromatografia líquida Intoxicações Dispersive liquid-liquid microextraction Liquid chromatography Intoxications CNPQ::CIENCIAS DA SAUDE::FARMACIA |
topic |
Microextração líquido-líquido dispersiva Benzodiazepínicos Plasma Cromatografia líquida Intoxicações Dispersive liquid-liquid microextraction Liquid chromatography Intoxications CNPQ::CIENCIAS DA SAUDE::FARMACIA |
description |
Drugs has been the main agents involved in intoxications in Brazil. According to the latest report of SINITOX, these agents summed up 20367 cases representing 26,79% of all agents involved. Among them, 9983 cases were suicide attempts. Psychotropic drugs of the benzodiazepine class were the agents with bigger representativity in this scenario. Among them, Clonazepam (Rivotril®) was involved in 1506 cases (1223 under intentional circumstance). Benzodiazepines possess pharmacological characteristics convenient for suicide attempts and as drug facilitated crimes. As these characteristics are not exclusive of this drug class, it is necessary a development of an analytical method capable of detect and quantify these drugs with adequate specificity and sensitivity to help in the doctor behavior and other health professionals in accidental or intentional intoxications. In this context, blood is considered a complex matrix that allows to determine the concentration of this xenobiotic which is still active in the organism, allowing to correlate its levels with the biological effects, besides being a sample of choice for toxicological analysis. However, extractive procedures are necessary as previous step to promote biological matrix clean up, avoiding equipment damage and to other inputs as concentrate the analytes of interest allowing the chromatographic system detection. Among the extractive techniques, the dispersive liquid-liquid microextraction (DLLME) stands out for being a quick, simple, low cost process adaptable in any laboratory and it was little explored for the purpose of toxicological analyses. The main objective of this research was the development for an analytical method for the benzodiazepine analysis in plasma aiming to improve the medical behavior in accidental or intentional intoxications from the Hospital Universitário de Santa Maria located in the Universidade Federal de Santa Maria (HUSMUFSM). The results revealed that the extractive procedure got optimized this way, from 500 μL of plasma, previously alkalized to pH 10, are added 0,013 g of sodium chloride, then 400 μL of chloroform and 700 μL of acetonitrile are injected simultaneously, the content is homogenized in vortex for 10 seconds, then it is placed under ultrasonic bath por 1 minute, then the tube is centrifuged by 10000 rpm/10 minutes, the organic phase is collected, dried and resurrected with 30 μL of mobile phase (water pH 9/methanol/acetonitrile, 63:19:18) and 20 μL in the HPLCDAD. In the analytical validation, according to UNODC and SWGTOX guides, it wasn’t possible to validate all the parameters, it is possible to apply this method only for screening. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-03-27 2021-06-21T17:03:40Z 2021-06-21T17:03:40Z |
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 |
http://repositorio.ufsm.br/handle/1/21163 |
dc.identifier.dark.fl_str_mv |
ark:/26339/0013000009vd5 |
url |
http://repositorio.ufsm.br/handle/1/21163 |
identifier_str_mv |
ark:/26339/0013000009vd5 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Análises Clínicas e Toxicológicas UFSM Programa de Pós-Graduação em Ciências Farmacêuticas Centro de Ciências da Saúde |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Análises Clínicas e Toxicológicas UFSM Programa de Pós-Graduação em Ciências Farmacêuticas Centro de Ciências da Saúde |
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 |
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1821325846940483584 |