Determinação de fraude de Leite com soro de leite pela análise de CMP e Pseudo-CMP por cromatografia líquida de alta eficiência em fase reversa com detecção por espectrometria de massa
Autor(a) principal: | |
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Data de Publicação: | 2008 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/2852 |
Resumo: | The main objective of this work was the development of a method of analysis by High Performance Liquid Chromatography with reversed phase and detection by mass spectrometry to determine the adulteration of milk with whey by detection caseinomacropeptídio (CMP), and determine the effect on the cultivation of milk with Pseudomonas fluorescens to verify the occurrence of pseudo-CMP or CMP produced by thermo-resistent enzymes of psychrotrophic bacteria. The milk samples were added of whey at concentrations of 0, 5, 10, 15 and 20% (v / v) and analyzed by High Performance Liquid Chromatography - Gélica Filtration (GF-HPLC) and High Performance Liquid Chromatography Reverse-phase with detection by Mass Spectrometry (HPLC- RP/MS). It was observed that there was a difference between the methods of analyses (p <0.05) and there was an effect of the whey concentration by two methods (p <0.05),that is, the increasing on the whey concentration also causes an increasing on the corresponding area of CMP by the two methods.The CMP was detected by HPLC-GF with retention time of around 5.7 minutes. When it was analyzed by HPLC- RP/MS, the CMP was detected at retention time around 19 minutes and characterized by ions of mass/charge 1697.4+4 and 2263.3+3. The samples of milk cultivated with Pseudomonas fluorescens were incubated on different days (0, 2, 4, 6 and 8) at 7 ºC, added in different concentrations of whey (0, 5, 10, 15 and 20% (v / v)) and analyzed by HPLC-FG and HPLC-RP/MS. On samples of milk added with Pseudomonas fluorescens and no addition of whey, analyzed by HPLC-FG, when incubated for up to 2 days, CMP was not found. Though, on samples of milk incubated for over 2 days, CMP was found; not deriving from the adulteration of milk with whey.The same samples of milk added to Pseudomonas fluorescens and no whey, analyzed by HPLC-RP/MS and incubated for up to 2 days did not present any increasing on the detection area of CMP. Though, on samples of milk incubated for over 2 days, CMP was found, characterized by ions of relation mass/charge 1697,4+4 and 2263,3+3 and other ions 1358+5, and the 1301,2+5. These latter ions derive from the hydrolysis of κ-casein by thermo-resistant enzymes of Pseudomonas fluorescens. The method of analyses by LAE- FG did not differentiate the CMP deriving from whey, from CMP and/or pseudo-CMP produced by proteasis of P. fluorescens. The analyses of CMP in the milk by CLAE-FR/EM were efficient to detect the adding of whey to the milk without proteolysis. Besides that, when the milk was proteolized by enzymes of P. fluorescens this method could indicate that there was proteolysis. Though, when the milk is proteolized by P. fluorescens, and when whey was added, there was no way of quantifying the CMP that was produced only by P. fluorescens and which was only due to the adding of whey. In such case, the result of the Index of CMP, as foreseen on IN 69/2006, is a great indicator of the intensity of proteolysis of milk protein, or then, from the fraudulent adding of milk and whey, when analyzed by CLAE-FR/EM, being then a great indicator of the quality of the milk. |
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Magalhães, Mirella Araújohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4713505H6Brandão, Sebastião César Cardosohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783541H2Andrade, Nélio José dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788281Y5Furtado, Mauro Mansurhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783310Z7Zacca, Jorge Jardimhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781655H0Fontes, Edimar Aparecida Filomenohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4792339T82015-03-26T13:13:13Z2009-07-072015-03-26T13:13:13Z2008-11-04MAGALHÃES, Mirella Araújo. Determining milk fraud with whey by analyses of CMP and Pseudo-CMP by High Performance Liquid Chromatography in reversed phase with detection by mass spectrometry. 2008. 74 f. Dissertação (Mestrado em Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos) - Universidade Federal de Viçosa, Viçosa, 2008.http://locus.ufv.br/handle/123456789/2852The main objective of this work was the development of a method of analysis by High Performance Liquid Chromatography with reversed phase and detection by mass spectrometry to determine the adulteration of milk with whey by detection caseinomacropeptídio (CMP), and determine the effect on the cultivation of milk with Pseudomonas fluorescens to verify the occurrence of pseudo-CMP or CMP produced by thermo-resistent enzymes of psychrotrophic bacteria. The milk samples were added of whey at concentrations of 0, 5, 10, 15 and 20% (v / v) and analyzed by High Performance Liquid Chromatography - Gélica Filtration (GF-HPLC) and High Performance Liquid Chromatography Reverse-phase with detection by Mass Spectrometry (HPLC- RP/MS). It was observed that there was a difference between the methods of analyses (p <0.05) and there was an effect of the whey concentration by two methods (p <0.05),that is, the increasing on the whey concentration also causes an increasing on the corresponding area of CMP by the two methods.The CMP was detected by HPLC-GF with retention time of around 5.7 minutes. When it was analyzed by HPLC- RP/MS, the CMP was detected at retention time around 19 minutes and characterized by ions of mass/charge 1697.4+4 and 2263.3+3. The samples of milk cultivated with Pseudomonas fluorescens were incubated on different days (0, 2, 4, 6 and 8) at 7 ºC, added in different concentrations of whey (0, 5, 10, 15 and 20% (v / v)) and analyzed by HPLC-FG and HPLC-RP/MS. On samples of milk added with Pseudomonas fluorescens and no addition of whey, analyzed by HPLC-FG, when incubated for up to 2 days, CMP was not found. Though, on samples of milk incubated for over 2 days, CMP was found; not deriving from the adulteration of milk with whey.The same samples of milk added to Pseudomonas fluorescens and no whey, analyzed by HPLC-RP/MS and incubated for up to 2 days did not present any increasing on the detection area of CMP. Though, on samples of milk incubated for over 2 days, CMP was found, characterized by ions of relation mass/charge 1697,4+4 and 2263,3+3 and other ions 1358+5, and the 1301,2+5. These latter ions derive from the hydrolysis of κ-casein by thermo-resistant enzymes of Pseudomonas fluorescens. The method of analyses by LAE- FG did not differentiate the CMP deriving from whey, from CMP and/or pseudo-CMP produced by proteasis of P. fluorescens. The analyses of CMP in the milk by CLAE-FR/EM were efficient to detect the adding of whey to the milk without proteolysis. Besides that, when the milk was proteolized by enzymes of P. fluorescens this method could indicate that there was proteolysis. Though, when the milk is proteolized by P. fluorescens, and when whey was added, there was no way of quantifying the CMP that was produced only by P. fluorescens and which was only due to the adding of whey. In such case, the result of the Index of CMP, as foreseen on IN 69/2006, is a great indicator of the intensity of proteolysis of milk protein, or then, from the fraudulent adding of milk and whey, when analyzed by CLAE-FR/EM, being then a great indicator of the quality of the milk.O objetivo principal desse trabalho foi o desenvolvimento de um método de análise laboratorial por cromatografia líquida de alta eficiência com Fase Reversa e detecção por Espectrometria de Massa para determinar a adulteração de leite com soro de leite através da detecção de caseinomacropeptídio (CMP), e determinar o efeito do cultivo do leite com Pseudomonas fluorescens para verificar a ocorrência de CMP ou pseudo-CMP produzido por enzimas termorresistentes de bactérias psicrotróficas. As amostras de leite foram adicionadas de soro de leite nas concentrações de 0; 5; 10; 15 e 20% (v/v) e analisadas por Cromatografia Liquida de Alta Eficiência Filtração Gélica (CLAE-FG) e Cromatografia Liquida de Alta Eficiência em Fase Reversa com detecção por Espectrometria de Massa (CLAE-FR/EM). Foi observado que houve diferença entre os métodos de análise (p<0,05) e houve efeito da concentração de soro de leite pelos dois métodos (p<0,05),ou seja, o aumento da concentração de soro acarreta um aumento da área correspondente ao CMP pelos dois métodos. O CMP foi detectado por CLAE-FG no tempo de retenção em torno de 5,7 minutos. Quando analisado por CLAE-FR/EM, o CMP foi detectado no tempo de retenção em torno de 19 minutos e caracterizado pelos íons de relação massa/ carga 1697,4+4 e 2263,3+3. As amostras de leite cultivadas com Pseudomonas fluorescens foram incubadas por diferentes dias (0, 2, 4, 6 e 8) a 7 ºC, adicionadas de diferentes concentrações de soro de leite (0; 5; 10; 15 e 20% (v/v)) e analisadas por CLAE-FG e CLAE-FR/EM. Nas amostras de leite adicionadas de Pseudomonas fluorescens e sem adição de soro de leite, analisadas por CLAE-FG, quando incubadas por até 2 dias, não foi verificado presença de CMP. Porém as amostras de leite incubadas por tempo acima de 2 dias foi verificada a presença de CMP, não proveniente da adulteração do leite com soro de leite. As mesmas amostras de leite adicionadas de Pseudomonas fluorescens e sem adição de soro, analisadas por CLAE-FR/EM, e incubadas por até 2 dias não apresentou aumento da área de detecção do CMP. Porém nas amostras de leite incubadas por tempo acima de 2 dias foi verificada a presença de CMP, caracterizado pelos íons de relação massa carga 1697,4+4 e 2263,3+3 e outros íons 1358+5, e o 1301,2+5. Estes últimos íons são oriundos da hidrólise da κ- caseína pelas enzimas termorresistentes de Pseudomonas fluorescens O método de análise por CLAE-FG não diferenciou o CMP proveniente do soro de leite, do CMP e/ou do pseudo- CMP produzido por proteases de P. fluorescens. A análise de CMP no leite por CLAE-FR/EM foi eficaz para detectar a adição de soro de leite ao leite sem proteólise. Além disso, quando o leite foi proteolizado por enzimas de P. fluorescens este método conseguiu indicar que houve proteólise. Porém, quando o leite estiver proteolisado por P. fluorescens, e estiver sido adicionado de soro, não tem como quantificar o CMP que foi produzido somente pela P. fluorescens e o que foi devido somente à adição de soro. Sendo assim, o resultado do Índice de CMP, como previsto na IN 69/2006, é um excelente indicador da intensidade de proteólise das proteínas do leite, ou então da adição fraudulenta do leite com soro de leite, quando analisado por CLAE-FR/EM, sendo um indicador eficiente da qualidade do leite.application/pdfporUniversidade Federal de ViçosaMestrado em Ciência e Tecnologia de AlimentosUFVBRCiência de Alimentos; Tecnologia de Alimentos; Engenharia de AlimentosCromatografia líquida de alta eficiênciaEspectrômetro de massaFraudeLeiteHigh performance liquid chromatographyMass spectrometryMilkFraudCNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOSDeterminação de fraude de Leite com soro de leite pela análise de CMP e Pseudo-CMP por cromatografia líquida de alta eficiência em fase reversa com detecção por espectrometria de massaDetermining milk fraud with whey by analyses of CMP and Pseudo-CMP by High Performance Liquid Chromatography in reversed phase with detection by mass spectrometryinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf686589https://locus.ufv.br//bitstream/123456789/2852/1/texto%20completo.pdf61ab5571f66b682dcf9918a19ecbc289MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain129064https://locus.ufv.br//bitstream/123456789/2852/2/texto%20completo.pdf.txtdf20a1df567ce301452169d995a5c8c3MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3868https://locus.ufv.br//bitstream/123456789/2852/3/texto%20completo.pdf.jpg1cb0ff350cb17734bbc9f1ed31cb3a6dMD53123456789/28522016-04-08 23:14:18.212oai:locus.ufv.br:123456789/2852Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-09T02:14:18LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Determinação de fraude de Leite com soro de leite pela análise de CMP e Pseudo-CMP por cromatografia líquida de alta eficiência em fase reversa com detecção por espectrometria de massa |
dc.title.alternative.eng.fl_str_mv |
Determining milk fraud with whey by analyses of CMP and Pseudo-CMP by High Performance Liquid Chromatography in reversed phase with detection by mass spectrometry |
title |
Determinação de fraude de Leite com soro de leite pela análise de CMP e Pseudo-CMP por cromatografia líquida de alta eficiência em fase reversa com detecção por espectrometria de massa |
spellingShingle |
Determinação de fraude de Leite com soro de leite pela análise de CMP e Pseudo-CMP por cromatografia líquida de alta eficiência em fase reversa com detecção por espectrometria de massa Magalhães, Mirella Araújo Cromatografia líquida de alta eficiência Espectrômetro de massa Fraude Leite High performance liquid chromatography Mass spectrometry Milk Fraud CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS |
title_short |
Determinação de fraude de Leite com soro de leite pela análise de CMP e Pseudo-CMP por cromatografia líquida de alta eficiência em fase reversa com detecção por espectrometria de massa |
title_full |
Determinação de fraude de Leite com soro de leite pela análise de CMP e Pseudo-CMP por cromatografia líquida de alta eficiência em fase reversa com detecção por espectrometria de massa |
title_fullStr |
Determinação de fraude de Leite com soro de leite pela análise de CMP e Pseudo-CMP por cromatografia líquida de alta eficiência em fase reversa com detecção por espectrometria de massa |
title_full_unstemmed |
Determinação de fraude de Leite com soro de leite pela análise de CMP e Pseudo-CMP por cromatografia líquida de alta eficiência em fase reversa com detecção por espectrometria de massa |
title_sort |
Determinação de fraude de Leite com soro de leite pela análise de CMP e Pseudo-CMP por cromatografia líquida de alta eficiência em fase reversa com detecção por espectrometria de massa |
author |
Magalhães, Mirella Araújo |
author_facet |
Magalhães, Mirella Araújo |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4713505H6 |
dc.contributor.author.fl_str_mv |
Magalhães, Mirella Araújo |
dc.contributor.advisor-co1.fl_str_mv |
Brandão, Sebastião César Cardoso |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783541H2 |
dc.contributor.advisor-co2.fl_str_mv |
Andrade, Nélio José de |
dc.contributor.advisor-co2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788281Y5 |
dc.contributor.advisor1.fl_str_mv |
Furtado, Mauro Mansur |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783310Z7 |
dc.contributor.referee1.fl_str_mv |
Zacca, Jorge Jardim |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781655H0 |
dc.contributor.referee2.fl_str_mv |
Fontes, Edimar Aparecida Filomeno |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4792339T8 |
contributor_str_mv |
Brandão, Sebastião César Cardoso Andrade, Nélio José de Furtado, Mauro Mansur Zacca, Jorge Jardim Fontes, Edimar Aparecida Filomeno |
dc.subject.por.fl_str_mv |
Cromatografia líquida de alta eficiência Espectrômetro de massa Fraude Leite |
topic |
Cromatografia líquida de alta eficiência Espectrômetro de massa Fraude Leite High performance liquid chromatography Mass spectrometry Milk Fraud CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS |
dc.subject.eng.fl_str_mv |
High performance liquid chromatography Mass spectrometry Milk Fraud |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS |
description |
The main objective of this work was the development of a method of analysis by High Performance Liquid Chromatography with reversed phase and detection by mass spectrometry to determine the adulteration of milk with whey by detection caseinomacropeptídio (CMP), and determine the effect on the cultivation of milk with Pseudomonas fluorescens to verify the occurrence of pseudo-CMP or CMP produced by thermo-resistent enzymes of psychrotrophic bacteria. The milk samples were added of whey at concentrations of 0, 5, 10, 15 and 20% (v / v) and analyzed by High Performance Liquid Chromatography - Gélica Filtration (GF-HPLC) and High Performance Liquid Chromatography Reverse-phase with detection by Mass Spectrometry (HPLC- RP/MS). It was observed that there was a difference between the methods of analyses (p <0.05) and there was an effect of the whey concentration by two methods (p <0.05),that is, the increasing on the whey concentration also causes an increasing on the corresponding area of CMP by the two methods.The CMP was detected by HPLC-GF with retention time of around 5.7 minutes. When it was analyzed by HPLC- RP/MS, the CMP was detected at retention time around 19 minutes and characterized by ions of mass/charge 1697.4+4 and 2263.3+3. The samples of milk cultivated with Pseudomonas fluorescens were incubated on different days (0, 2, 4, 6 and 8) at 7 ºC, added in different concentrations of whey (0, 5, 10, 15 and 20% (v / v)) and analyzed by HPLC-FG and HPLC-RP/MS. On samples of milk added with Pseudomonas fluorescens and no addition of whey, analyzed by HPLC-FG, when incubated for up to 2 days, CMP was not found. Though, on samples of milk incubated for over 2 days, CMP was found; not deriving from the adulteration of milk with whey.The same samples of milk added to Pseudomonas fluorescens and no whey, analyzed by HPLC-RP/MS and incubated for up to 2 days did not present any increasing on the detection area of CMP. Though, on samples of milk incubated for over 2 days, CMP was found, characterized by ions of relation mass/charge 1697,4+4 and 2263,3+3 and other ions 1358+5, and the 1301,2+5. These latter ions derive from the hydrolysis of κ-casein by thermo-resistant enzymes of Pseudomonas fluorescens. The method of analyses by LAE- FG did not differentiate the CMP deriving from whey, from CMP and/or pseudo-CMP produced by proteasis of P. fluorescens. The analyses of CMP in the milk by CLAE-FR/EM were efficient to detect the adding of whey to the milk without proteolysis. Besides that, when the milk was proteolized by enzymes of P. fluorescens this method could indicate that there was proteolysis. Though, when the milk is proteolized by P. fluorescens, and when whey was added, there was no way of quantifying the CMP that was produced only by P. fluorescens and which was only due to the adding of whey. In such case, the result of the Index of CMP, as foreseen on IN 69/2006, is a great indicator of the intensity of proteolysis of milk protein, or then, from the fraudulent adding of milk and whey, when analyzed by CLAE-FR/EM, being then a great indicator of the quality of the milk. |
publishDate |
2008 |
dc.date.issued.fl_str_mv |
2008-11-04 |
dc.date.available.fl_str_mv |
2009-07-07 2015-03-26T13:13:13Z |
dc.date.accessioned.fl_str_mv |
2015-03-26T13:13:13Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
MAGALHÃES, Mirella Araújo. Determining milk fraud with whey by analyses of CMP and Pseudo-CMP by High Performance Liquid Chromatography in reversed phase with detection by mass spectrometry. 2008. 74 f. Dissertação (Mestrado em Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos) - Universidade Federal de Viçosa, Viçosa, 2008. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/2852 |
identifier_str_mv |
MAGALHÃES, Mirella Araújo. Determining milk fraud with whey by analyses of CMP and Pseudo-CMP by High Performance Liquid Chromatography in reversed phase with detection by mass spectrometry. 2008. 74 f. Dissertação (Mestrado em Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos) - Universidade Federal de Viçosa, Viçosa, 2008. |
url |
http://locus.ufv.br/handle/123456789/2852 |
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Universidade Federal de Viçosa |
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Mestrado em Ciência e Tecnologia de Alimentos |
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UFV |
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BR |
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Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos |
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Universidade Federal de Viçosa |
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