Avaliação de métodos analíticos para estudos em material biológico de ratos submetidos ao tratamento com nanotubos de carbono

Detalhes bibliográficos
Ano de defesa: 2020
Autor(a) principal: Krzyzaniak, Sindy Raquel lattes
Orientador(a): Mello, Paola de Azevedo lattes
Banca de defesa: Pozebon, Dirce, Mortari, Sergio Roberto, Müller, Edson Irineu, Chitolina, Maria Rosa
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Santa Maria
Centro de Ciências Naturais e Exatas
Programa de Pós-Graduação: Programa de Pós-Graduação em Química
Departamento: Química
País: Brasil
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: http://repositorio.ufsm.br/handle/1/23091
Resumo: In this work, methods for the analysis of tissues, fluids and furs of rats submitted to the treatment with carbon nanotubes (CNTs) were evaluated, allowing to identify where they are concentrated, indirectly, by the determination of metal contaminants from CNTs. Contaminants determination was carried out by inductively coupled plasma optical emission spectrometry (ICP-OES) and/or inductively coupled plasma mass spectrometry (ICP-MS). Also, the distribution of inorganic contaminants in the tissues of animals was evaluated through bio-imaging by laser ablation (LA) combined to ICP-MS (LA-ICP-MS). Initially, it was necessary to obtain the tissues (spleen, brain, heart, liver, pancreas, lung and kidney), fluids (feces and blood) and furs of animals submitted to the treatment with CNTs, as suggested by the Comitê de Ética Animal of UFSM, with the study of acute oral toxicity and chronic toxicity. In this study, a functionalized CNT with hydroxyl groups (CNT-OH) was used. In the acute oral toxicity test, CNTs shown to be non-toxic when administered to rats at the maximum dose of 2000 mg kg-1 per body weight. According to the histological analyzes of the liver, doses of 300 mg kg-1 and 2000 mg kg-1 caused small hepatic lesions. Because of this, 1 mg kg-1, 10 mg kg-1 and 100 mg kg-1 of CNT were administered in the chronic toxicity study for 31 days. Oral administration of CNTs did not cause significant changes in biochemical parameters and at the end of the treatment, the rats were euthanized and the organs, fluids, furs and feces of the animals were collected. The need to evaluate tissues contaminated with CNTs required the development of a method that was efficient for the decomposition of this matrix, a challenge in terms of sample preparation, to avoid results underestimated by the inefficient digestion of the CNT into the tissue. In this purpose, the decomposition method in open system with digestion heating block was evaluated, as well as a method based on microwave-assisted wet digestion using a pressurized digestion cavity (MAWD-PDC). Sample preparation using the digestion block was considered unsuitable once it does not allowed the decomposition of CNTs. For MAWD-PDC, the influence of the mass of animal tissue to be decomposed and the efficiency for the digestion of CNTs contained in the tissues were investigated. The accuracy of the method was evaluated with the decomposition of a reference sample of bovine liver, added to samples of bovine liver and chicken, used to simulate the tissues. Suitable recoveries (90 to 127%) were observed. Results obtained using 275 mg of CNT for decomposition by MAWD-PDC, presented no difference (t-student test) when compared with reference values. After optimization, the MAWD-PDC was considered suitable for the decomposition of tissues, fluids and furs of animals submitted to the treatment with CNTs using 6 mL of concentrated HNO3 and 0.5 mL of 30% H2O2 as a decomposition solution and a program consisting of i) 25 min at 180 °C (10 min ramp) and ii) 40 min at 270 °C (5 min ramp). In general, differences were observed between the results obtained for Ni in the liver, heart, lung and feces of the animals in the groups treated with CNTs in relation to the control group. The bio-distribution analysis by LA-ICP-MS in the spleen, liver, lung and kidney of the animals from the control group and those from the 100 mg kg-1 CNT treated group was performed. Bio-imaging of the distribution of elements considered macronutrients, such as Ca and Mg, and micronutrients as Cu, Fe, Mn and Zn, were obtained. Differences in the distribution of some elements in the kidney, spleen, and lung were observed. In view of the results obtained using MAWD-PDC and the determination of metals by ICP-OES and/or ICP-MS and the bio-distribution analysis by LA-ICP-MS, it can be inferred that both investigated strategies were promising and can be used contributing to the assessment of the possible toxic effects of CNTs.
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spelling 2021-12-01T14:39:49Z2021-12-01T14:39:49Z2020-05-27http://repositorio.ufsm.br/handle/1/23091In this work, methods for the analysis of tissues, fluids and furs of rats submitted to the treatment with carbon nanotubes (CNTs) were evaluated, allowing to identify where they are concentrated, indirectly, by the determination of metal contaminants from CNTs. Contaminants determination was carried out by inductively coupled plasma optical emission spectrometry (ICP-OES) and/or inductively coupled plasma mass spectrometry (ICP-MS). Also, the distribution of inorganic contaminants in the tissues of animals was evaluated through bio-imaging by laser ablation (LA) combined to ICP-MS (LA-ICP-MS). Initially, it was necessary to obtain the tissues (spleen, brain, heart, liver, pancreas, lung and kidney), fluids (feces and blood) and furs of animals submitted to the treatment with CNTs, as suggested by the Comitê de Ética Animal of UFSM, with the study of acute oral toxicity and chronic toxicity. In this study, a functionalized CNT with hydroxyl groups (CNT-OH) was used. In the acute oral toxicity test, CNTs shown to be non-toxic when administered to rats at the maximum dose of 2000 mg kg-1 per body weight. According to the histological analyzes of the liver, doses of 300 mg kg-1 and 2000 mg kg-1 caused small hepatic lesions. Because of this, 1 mg kg-1, 10 mg kg-1 and 100 mg kg-1 of CNT were administered in the chronic toxicity study for 31 days. Oral administration of CNTs did not cause significant changes in biochemical parameters and at the end of the treatment, the rats were euthanized and the organs, fluids, furs and feces of the animals were collected. The need to evaluate tissues contaminated with CNTs required the development of a method that was efficient for the decomposition of this matrix, a challenge in terms of sample preparation, to avoid results underestimated by the inefficient digestion of the CNT into the tissue. In this purpose, the decomposition method in open system with digestion heating block was evaluated, as well as a method based on microwave-assisted wet digestion using a pressurized digestion cavity (MAWD-PDC). Sample preparation using the digestion block was considered unsuitable once it does not allowed the decomposition of CNTs. For MAWD-PDC, the influence of the mass of animal tissue to be decomposed and the efficiency for the digestion of CNTs contained in the tissues were investigated. The accuracy of the method was evaluated with the decomposition of a reference sample of bovine liver, added to samples of bovine liver and chicken, used to simulate the tissues. Suitable recoveries (90 to 127%) were observed. Results obtained using 275 mg of CNT for decomposition by MAWD-PDC, presented no difference (t-student test) when compared with reference values. After optimization, the MAWD-PDC was considered suitable for the decomposition of tissues, fluids and furs of animals submitted to the treatment with CNTs using 6 mL of concentrated HNO3 and 0.5 mL of 30% H2O2 as a decomposition solution and a program consisting of i) 25 min at 180 °C (10 min ramp) and ii) 40 min at 270 °C (5 min ramp). In general, differences were observed between the results obtained for Ni in the liver, heart, lung and feces of the animals in the groups treated with CNTs in relation to the control group. The bio-distribution analysis by LA-ICP-MS in the spleen, liver, lung and kidney of the animals from the control group and those from the 100 mg kg-1 CNT treated group was performed. Bio-imaging of the distribution of elements considered macronutrients, such as Ca and Mg, and micronutrients as Cu, Fe, Mn and Zn, were obtained. Differences in the distribution of some elements in the kidney, spleen, and lung were observed. In view of the results obtained using MAWD-PDC and the determination of metals by ICP-OES and/or ICP-MS and the bio-distribution analysis by LA-ICP-MS, it can be inferred that both investigated strategies were promising and can be used contributing to the assessment of the possible toxic effects of CNTs.Neste trabalho foram avaliados métodos para a análise de tecidos, fluidos e pelos de ratos submetidos a tratamentos com nanotubos de carbono (CNTs), possibilitando identificar onde estes se concentram, de maneira indireta, pela determinação de metais contaminantes dos CNTs. A determinação dos contaminantes nos tecidos, fluidos e pelos foi feita por espectrometria de emissão óptica com plasma indutivamente acoplado (ICP-OES) e/ou espectrometria de massa com plasma indutivamente acoplado (ICP-MS). Além disso, foi feita a avaliação da distribuição dos contaminantes inorgânicos nos tecidos dos animais, através da obtenção de bioimagens, utilizando a técnica de ablação a laser (LA) combinada com ICP-MS (LA-ICP-MS). Inicialmente, foi necessário obter os tecidos (baço, cérebro, coração, fígado, pâncreas, pulmão e rim), fluidos (fezes e sangue) e pelos de animais submetidos a tratamentos com CNTs, conforme sugestão do Comitê de Ética Animal da UFSM, com a realização do estudo de toxicidade oral aguda e de toxicidade crônica. Nesse estudo foi usado um CNT funcionalizado com grupos hidroxila (CNT-OH). No teste de toxicidade oral aguda, os CNTs mostraram não ser tóxicos quando administrados em ratas na dose máxima de 2000 mg kg-1 de peso corpóreo. De acordo com a análise histológica do fígado, as doses de 300 e 2000 mg kg-1 provocaram pequenas lesões hepáticas. Devido a isto, no estudo de toxicidade crônica por 31 dias, foi feita a administração de 1 mg kg-1, 10 mg kg-1 e 100 mg kg-1 de CNT. A administração oral de CNTs não provocou alterações significativas em parâmetros bioquímicos e no final do tratamento os ratos foram eutanasiados e os orgãos, fluidos, pelos e fezes dos animais foram coletados. A necessidade de avaliar tecidos contaminados com CNTs exigiu o desenvolvimento de um método que apresentasse eficiência para a decomposição dessa matriz, um desafio em termos de preparo de amostras, para evitar resultados subestimados pela decomposição ineficiente do CNT presente no tecido. Para tal, foi avaliado o método de decomposição em sistema aberto com aquecimento em bloco digestor, bem como um método baseado na digestão por via úmida assistida por micro-ondas em uma cavidade de digestão pressurizada (MAWD-PDC). O preparo da amostra usando bloco digestor foi considerado inadequado haja vista que não possibilitou a decomposição dos CNTs. Na avaliação da MAWD-PDC foi investigada a influência da massa de tecido animal a ser decomposta e a eficiência de decomposição dos CNTs contidos nos tecidos. A exatidão do método foi avaliada com a decomposição de uma amostra de referência de fígado bovino, adicionada à amostras de fígado bovino e frango, usadas para simular os tecidos e foram observadas recuperações adequadas (90 a 127%). Os resultados obtidos utilizando 275 mg de CNT não apresentaram diferença (teste t-student) quando comparados a valores de referência. Após estas avaliações, a MAWD-PCD foi considerada adequada para a decomposição de tecidos, fluidos e pelos dos animais submetidos a tratamentos com CNTs. empregando 6 mL de HNO3 concentrado e 0,5 mL de H2O2 30% como solução de decomposição e um programa constituído de i) 25 min a 180 °C (10 min de rampa) e ii) 40 min a 270 °C (5 min de rampa). De maneira geral, foram observadas diferenças entre os resultados obtidos para Ni no fígado, coração, pulmão e nas fezes dos animais pertencentes aos grupos tratados com CNTs em relação ao grupo controle. A análise de biodistribuição por LA-ICP-MS foi feita no baço, fígado, pulmão e rim dos animais do grupo controle e daqueles pertencentes ao grupo tratado com 100 mg kg-1 de CNT. Foram obtidas as bioimagens da distribuição dos elementos considerados macronutrientes, como o Ca e Mg, e micronutrientes como o Cu, Fe, Mn e Zn, sendo observada diferença na distribuição de alguns elementos no rim, baço e pulmão. Tendo em vista os resultados obtidos usando MAWD-PDC e determinação de metais por ICP-OES e/ou ICP-MS e considerando as análises de biodistribuição por LA-ICP-MS, pode-se inferir que ambas as estratégias investigadas foram promissoras e podem ser utilizadas, contribuindo para a avaliação dos possíveis efeitos tóxicos dos CNTs.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqporUniversidade Federal de Santa MariaCentro de Ciências Naturais e ExatasPrograma de Pós-Graduação em QuímicaUFSMBrasilQuímicaAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessNanotubos de carbonoContaminantes inorgânicosAnálise de biodistribuiçãoCarbon nanotubesInorganic contaminantsBio-distribution analysisCNPQ::CIENCIAS EXATAS E DA TERRA::QUIMICAAvaliação de métodos analíticos para estudos em material biológico de ratos submetidos ao tratamento com nanotubos de carbonoEvaluation of analytical methods for studies on rat biological material submitted to treatment with carbon nanotubes.info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisMello, Paola de Azevedohttp://lattes.cnpq.br/2189500441942469Pozebon, DirceMortari, Sergio RobertoMüller, Edson IrineuChitolina, Maria Rosahttp://lattes.cnpq.br/9088666879040816Krzyzaniak, Sindy Raquel1006000000006006006006006000128b934-4b3f-43d6-8526-fb278da0f673a866c1af-fc31-48d2-99f7-dada20cc9415a10aee69-497f-4257-96b7-c8a42f2d96833c621a6d-38be-4e61-b644-40b0cc92afcb5340e61b-ef9c-45e3-9754-7c9649711f614c1ed42a-03f5-4ced-bf2d-b4bdcf46311creponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALTES_PPGQUIMICA_2020_KRZYZANIAK_SINDY.pdfTES_PPGQUIMICA_2020_KRZYZANIAK_SINDY.pdfTese de Doutoradoapplication/pdf4560871http://repositorio.ufsm.br/bitstream/1/23091/1/TES_PPGQUIMICA_2020_KRZYZANIAK_SINDY.pdf3ae7c88a88934d10893e8afc014e9533MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; 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dc.title.por.fl_str_mv Avaliação de métodos analíticos para estudos em material biológico de ratos submetidos ao tratamento com nanotubos de carbono
dc.title.alternative.eng.fl_str_mv Evaluation of analytical methods for studies on rat biological material submitted to treatment with carbon nanotubes.
title Avaliação de métodos analíticos para estudos em material biológico de ratos submetidos ao tratamento com nanotubos de carbono
spellingShingle Avaliação de métodos analíticos para estudos em material biológico de ratos submetidos ao tratamento com nanotubos de carbono
Krzyzaniak, Sindy Raquel
Nanotubos de carbono
Contaminantes inorgânicos
Análise de biodistribuição
Carbon nanotubes
Inorganic contaminants
Bio-distribution analysis
CNPQ::CIENCIAS EXATAS E DA TERRA::QUIMICA
title_short Avaliação de métodos analíticos para estudos em material biológico de ratos submetidos ao tratamento com nanotubos de carbono
title_full Avaliação de métodos analíticos para estudos em material biológico de ratos submetidos ao tratamento com nanotubos de carbono
title_fullStr Avaliação de métodos analíticos para estudos em material biológico de ratos submetidos ao tratamento com nanotubos de carbono
title_full_unstemmed Avaliação de métodos analíticos para estudos em material biológico de ratos submetidos ao tratamento com nanotubos de carbono
title_sort Avaliação de métodos analíticos para estudos em material biológico de ratos submetidos ao tratamento com nanotubos de carbono
author Krzyzaniak, Sindy Raquel
author_facet Krzyzaniak, Sindy Raquel
author_role author
dc.contributor.advisor1.fl_str_mv Mello, Paola de Azevedo
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/2189500441942469
dc.contributor.referee1.fl_str_mv Pozebon, Dirce
dc.contributor.referee2.fl_str_mv Mortari, Sergio Roberto
dc.contributor.referee3.fl_str_mv Müller, Edson Irineu
dc.contributor.referee4.fl_str_mv Chitolina, Maria Rosa
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/9088666879040816
dc.contributor.author.fl_str_mv Krzyzaniak, Sindy Raquel
contributor_str_mv Mello, Paola de Azevedo
Pozebon, Dirce
Mortari, Sergio Roberto
Müller, Edson Irineu
Chitolina, Maria Rosa
dc.subject.por.fl_str_mv Nanotubos de carbono
Contaminantes inorgânicos
Análise de biodistribuição
topic Nanotubos de carbono
Contaminantes inorgânicos
Análise de biodistribuição
Carbon nanotubes
Inorganic contaminants
Bio-distribution analysis
CNPQ::CIENCIAS EXATAS E DA TERRA::QUIMICA
dc.subject.eng.fl_str_mv Carbon nanotubes
Inorganic contaminants
Bio-distribution analysis
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS EXATAS E DA TERRA::QUIMICA
description In this work, methods for the analysis of tissues, fluids and furs of rats submitted to the treatment with carbon nanotubes (CNTs) were evaluated, allowing to identify where they are concentrated, indirectly, by the determination of metal contaminants from CNTs. Contaminants determination was carried out by inductively coupled plasma optical emission spectrometry (ICP-OES) and/or inductively coupled plasma mass spectrometry (ICP-MS). Also, the distribution of inorganic contaminants in the tissues of animals was evaluated through bio-imaging by laser ablation (LA) combined to ICP-MS (LA-ICP-MS). Initially, it was necessary to obtain the tissues (spleen, brain, heart, liver, pancreas, lung and kidney), fluids (feces and blood) and furs of animals submitted to the treatment with CNTs, as suggested by the Comitê de Ética Animal of UFSM, with the study of acute oral toxicity and chronic toxicity. In this study, a functionalized CNT with hydroxyl groups (CNT-OH) was used. In the acute oral toxicity test, CNTs shown to be non-toxic when administered to rats at the maximum dose of 2000 mg kg-1 per body weight. According to the histological analyzes of the liver, doses of 300 mg kg-1 and 2000 mg kg-1 caused small hepatic lesions. Because of this, 1 mg kg-1, 10 mg kg-1 and 100 mg kg-1 of CNT were administered in the chronic toxicity study for 31 days. Oral administration of CNTs did not cause significant changes in biochemical parameters and at the end of the treatment, the rats were euthanized and the organs, fluids, furs and feces of the animals were collected. The need to evaluate tissues contaminated with CNTs required the development of a method that was efficient for the decomposition of this matrix, a challenge in terms of sample preparation, to avoid results underestimated by the inefficient digestion of the CNT into the tissue. In this purpose, the decomposition method in open system with digestion heating block was evaluated, as well as a method based on microwave-assisted wet digestion using a pressurized digestion cavity (MAWD-PDC). Sample preparation using the digestion block was considered unsuitable once it does not allowed the decomposition of CNTs. For MAWD-PDC, the influence of the mass of animal tissue to be decomposed and the efficiency for the digestion of CNTs contained in the tissues were investigated. The accuracy of the method was evaluated with the decomposition of a reference sample of bovine liver, added to samples of bovine liver and chicken, used to simulate the tissues. Suitable recoveries (90 to 127%) were observed. Results obtained using 275 mg of CNT for decomposition by MAWD-PDC, presented no difference (t-student test) when compared with reference values. After optimization, the MAWD-PDC was considered suitable for the decomposition of tissues, fluids and furs of animals submitted to the treatment with CNTs using 6 mL of concentrated HNO3 and 0.5 mL of 30% H2O2 as a decomposition solution and a program consisting of i) 25 min at 180 °C (10 min ramp) and ii) 40 min at 270 °C (5 min ramp). In general, differences were observed between the results obtained for Ni in the liver, heart, lung and feces of the animals in the groups treated with CNTs in relation to the control group. The bio-distribution analysis by LA-ICP-MS in the spleen, liver, lung and kidney of the animals from the control group and those from the 100 mg kg-1 CNT treated group was performed. Bio-imaging of the distribution of elements considered macronutrients, such as Ca and Mg, and micronutrients as Cu, Fe, Mn and Zn, were obtained. Differences in the distribution of some elements in the kidney, spleen, and lung were observed. In view of the results obtained using MAWD-PDC and the determination of metals by ICP-OES and/or ICP-MS and the bio-distribution analysis by LA-ICP-MS, it can be inferred that both investigated strategies were promising and can be used contributing to the assessment of the possible toxic effects of CNTs.
publishDate 2020
dc.date.issued.fl_str_mv 2020-05-27
dc.date.accessioned.fl_str_mv 2021-12-01T14:39:49Z
dc.date.available.fl_str_mv 2021-12-01T14:39:49Z
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dc.identifier.uri.fl_str_mv http://repositorio.ufsm.br/handle/1/23091
url http://repositorio.ufsm.br/handle/1/23091
dc.language.iso.fl_str_mv por
language por
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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.publisher.none.fl_str_mv Universidade Federal de Santa Maria
Centro de Ciências Naturais e Exatas
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Química
dc.publisher.initials.fl_str_mv UFSM
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Química
publisher.none.fl_str_mv Universidade Federal de Santa Maria
Centro de Ciências Naturais e Exatas
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instname_str Universidade Federal de Santa Maria (UFSM)
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