Estudo da degradação e potencial antigênico de proteínas do ovo no processamento de biscoitos semidoces e validação de método imunoenzimático

Detalhes bibliográficos
Autor(a) principal: Pedro Paulo Borges dos Santos
Data de Publicação: 2017
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFMG
Texto Completo: http://hdl.handle.net/1843/BUOS-BATFN3
Resumo: The prevalence of food allergy is about 5 % in children and 3 % to 4 % in teenagers and adults. The food allergens can cause adverse effects even in small amounts. Among them, egg proteins are highlighted by high prevalence, being susceptible to changes in processing. Few existing methods for food allergen determination comply with the performance parameters required for validation. Therefore, the aims of this work was study the degradation of egg white proteins in semi-sweet biscuits, under different processing conditions, and validate an immunoenzymatic kit for this scope. Formulations containing 0,022 % of egg were baked in different times (5, 10, 15, 20, 25 and 30 minutes) and temperatures (150, 180 and 210 °C), in a factorial design, and were analyzed by the kit. In the single-laboratory validation, two different analytical batches of semi-sweet biscuits spiked with ovalbumin standard in nine concentration levels (0.125, 0.185, 0.25, 0.5, 1.0, 3.0, 4.5, 9 and 13.5 mg/kg) in 10 replicates, plus the blank, were analyzed. A significant reduction (p <0.05) in the egg protein content was observed in baked biscuits. With 25 minutes of baking, were evidenced reductions of 83.1; 92.6 and 100% for the temperatures 150, 180 and 210 ° C, respectively, indicating influence of the processing on the antigenic potential. The results of the quantitative approach of validation showed that the performance criteria were not complied. In the qualitative approach, 100 % of sensitivity and reliability rates were found for the blank samples. In the levels 0.125; 0.185 and 0.25 mg/kg the sensitivity and reliability rates were 0; 90 and 80 %, respectively. From the level 0.5 mg/kg, the estimated values of these rates were 100 %, demonstrating sensitivity of the method. The accordance ranged from 0.5 to 1.0 and concordance from 0.7 to 1.0. For the concentrations of 0; 0.125 and over 0.5 mg/kg the concordance reaches maximum values, showing standardization of the method. The detection limit was established as 0.2 mg/kg. The studied kit presented selectivity in the presence of the other allergenic protein beta-lactoglobulin. Thus, the baking conditions were evidenced as critical for the antigenicity of egg white proteins and the studied kit was considered appropriate for the detection of egg white proteins in semi-sweet biscuits, characterizing an important tool for the implementation of the control of food allergens labeling.
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spelling Estudo da degradação e potencial antigênico de proteínas do ovo no processamento de biscoitos semidoces e validação de método imunoenzimáticoValidação de MétodosDegradação de ProteínasProteínas do OvoAlergia AlimentarELISAValidação de métodoTeste imunoenzimáticoAlimentos AnáliseAlergia a alimentosThe prevalence of food allergy is about 5 % in children and 3 % to 4 % in teenagers and adults. The food allergens can cause adverse effects even in small amounts. Among them, egg proteins are highlighted by high prevalence, being susceptible to changes in processing. Few existing methods for food allergen determination comply with the performance parameters required for validation. Therefore, the aims of this work was study the degradation of egg white proteins in semi-sweet biscuits, under different processing conditions, and validate an immunoenzymatic kit for this scope. Formulations containing 0,022 % of egg were baked in different times (5, 10, 15, 20, 25 and 30 minutes) and temperatures (150, 180 and 210 °C), in a factorial design, and were analyzed by the kit. In the single-laboratory validation, two different analytical batches of semi-sweet biscuits spiked with ovalbumin standard in nine concentration levels (0.125, 0.185, 0.25, 0.5, 1.0, 3.0, 4.5, 9 and 13.5 mg/kg) in 10 replicates, plus the blank, were analyzed. A significant reduction (p <0.05) in the egg protein content was observed in baked biscuits. With 25 minutes of baking, were evidenced reductions of 83.1; 92.6 and 100% for the temperatures 150, 180 and 210 ° C, respectively, indicating influence of the processing on the antigenic potential. The results of the quantitative approach of validation showed that the performance criteria were not complied. In the qualitative approach, 100 % of sensitivity and reliability rates were found for the blank samples. In the levels 0.125; 0.185 and 0.25 mg/kg the sensitivity and reliability rates were 0; 90 and 80 %, respectively. From the level 0.5 mg/kg, the estimated values of these rates were 100 %, demonstrating sensitivity of the method. The accordance ranged from 0.5 to 1.0 and concordance from 0.7 to 1.0. For the concentrations of 0; 0.125 and over 0.5 mg/kg the concordance reaches maximum values, showing standardization of the method. The detection limit was established as 0.2 mg/kg. The studied kit presented selectivity in the presence of the other allergenic protein beta-lactoglobulin. Thus, the baking conditions were evidenced as critical for the antigenicity of egg white proteins and the studied kit was considered appropriate for the detection of egg white proteins in semi-sweet biscuits, characterizing an important tool for the implementation of the control of food allergens labeling.A alergia alimentar possui uma prevalência de aproximadamente 5 % em crianças e de 3 a 4 % em adolescentes e adultos. Os alimentos alergênicos podem causar efeitos adversos, mesmo em pequenas concentrações. Dentre eles, as proteínas do ovo se destacam pela prevalência elevada, sendo susceptíveis a alterações no processamento. Poucos métodos existentes para determinação de alergênicos em alimentos cumprem com os parâmetros de desempenho necessários para validação. Portanto, o objetivo desse trabalho foi estudar a degradação de proteínas da clara do ovo em biscoitos semidoces, submetidos a diferentes condições de processamento, e validar um kit imunoenzimático para o referido escopo analítico. Foram preparadas formulações com 0,022 % de ovo as quais foram assadas por diferentes tempos (5, 10, 15, 20, 25 e 30 minutos) e temperaturas (150; 180 e 210 °C), em um delineamento fatorial, as quais foram analisadas pelo kit. Na validação intralaboratorial, foram analisados, em duas baterias analíticas distintas, biscoitos semidoces incorporados de padrão de ovoalbumina, em nove níveis de concentração (0,125; 0,185; 0,25; 0,5; 1,0; 3,0; 4,5; 9 e 13,5 mg/kg) e 10 replicatas, mais o branco. Foi observada redução significativa (p < 0,05) do teor de proteínas da clara nos biscoitos assados. Com 25 minutos de assamento, foram evidenciadas quedas de 83,1; 92,6 e 100 % para as temperaturas 150, 180 e 210 °C, respectivamente, indicando influência do processamento no potencial antigênico. Na análise dos resultados da validação, sob uma abordagem quantitativa, percebeu-se que os critérios de desempenho não foram atendidos. Pela abordagem qualitativa, foi encontrado 100 % de taxa de seletividade e confiabilidade para as amostras brancas. Nos níveis 0,125; 0,185 e 0,25 mg/kg, as taxas de sensibilidade e confiabilidade encontradas foram de 0; 90 e 80 %, respectivamente. A partir da concentração 0,5 mg/kg, os valores estimados para essas taxas foram de 100 %, demonstrando sensibilidade do método. A acordância variou de 0,5 a 1,0 e a concordância de 0,7 a 1,0. Nas concentrações 0; 0,125 e acima de 0,5 mg/kg, a concordância atingiu valores máximos, o que mostrou padronização adequada do método. O limite de detecção estabelecido foi de 0,2 mg/kg. O kit estudado apresentou, ainda, seletividade em relação à outra proteína alergênica, a betalactoglobulina. Desta forma, as condições de assamento foram evidenciadas como determinantes da antigenicidade das proteínas da clara e o kit estudado foi considerado apropriado para a detecção de proteínas da clara em biscoitos, caracterizando-se uma importante ferramenta para o controle da rotulagem alérgenos em alimentos.Universidade Federal de Minas GeraisUFMGScheilla Vitorino de Souza FerreiraPedro Paulo Borges dos Santos2019-08-10T22:22:14Z2019-08-10T22:22:14Z2017-02-23info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfhttp://hdl.handle.net/1843/BUOS-BATFN3info:eu-repo/semantics/openAccessporreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMG2021-04-08T21:09:17Zoai:repositorio.ufmg.br:1843/BUOS-BATFN3Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oairepositorio@ufmg.bropendoar:2021-04-08T21:09:17Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
dc.title.none.fl_str_mv Estudo da degradação e potencial antigênico de proteínas do ovo no processamento de biscoitos semidoces e validação de método imunoenzimático
title Estudo da degradação e potencial antigênico de proteínas do ovo no processamento de biscoitos semidoces e validação de método imunoenzimático
spellingShingle Estudo da degradação e potencial antigênico de proteínas do ovo no processamento de biscoitos semidoces e validação de método imunoenzimático
Pedro Paulo Borges dos Santos
Validação de Métodos
Degradação de Proteínas
Proteínas do Ovo
Alergia Alimentar
ELISA
Validação de método
Teste imunoenzimático
Alimentos Análise
Alergia a alimentos
title_short Estudo da degradação e potencial antigênico de proteínas do ovo no processamento de biscoitos semidoces e validação de método imunoenzimático
title_full Estudo da degradação e potencial antigênico de proteínas do ovo no processamento de biscoitos semidoces e validação de método imunoenzimático
title_fullStr Estudo da degradação e potencial antigênico de proteínas do ovo no processamento de biscoitos semidoces e validação de método imunoenzimático
title_full_unstemmed Estudo da degradação e potencial antigênico de proteínas do ovo no processamento de biscoitos semidoces e validação de método imunoenzimático
title_sort Estudo da degradação e potencial antigênico de proteínas do ovo no processamento de biscoitos semidoces e validação de método imunoenzimático
author Pedro Paulo Borges dos Santos
author_facet Pedro Paulo Borges dos Santos
author_role author
dc.contributor.none.fl_str_mv Scheilla Vitorino de Souza Ferreira
dc.contributor.author.fl_str_mv Pedro Paulo Borges dos Santos
dc.subject.por.fl_str_mv Validação de Métodos
Degradação de Proteínas
Proteínas do Ovo
Alergia Alimentar
ELISA
Validação de método
Teste imunoenzimático
Alimentos Análise
Alergia a alimentos
topic Validação de Métodos
Degradação de Proteínas
Proteínas do Ovo
Alergia Alimentar
ELISA
Validação de método
Teste imunoenzimático
Alimentos Análise
Alergia a alimentos
description The prevalence of food allergy is about 5 % in children and 3 % to 4 % in teenagers and adults. The food allergens can cause adverse effects even in small amounts. Among them, egg proteins are highlighted by high prevalence, being susceptible to changes in processing. Few existing methods for food allergen determination comply with the performance parameters required for validation. Therefore, the aims of this work was study the degradation of egg white proteins in semi-sweet biscuits, under different processing conditions, and validate an immunoenzymatic kit for this scope. Formulations containing 0,022 % of egg were baked in different times (5, 10, 15, 20, 25 and 30 minutes) and temperatures (150, 180 and 210 °C), in a factorial design, and were analyzed by the kit. In the single-laboratory validation, two different analytical batches of semi-sweet biscuits spiked with ovalbumin standard in nine concentration levels (0.125, 0.185, 0.25, 0.5, 1.0, 3.0, 4.5, 9 and 13.5 mg/kg) in 10 replicates, plus the blank, were analyzed. A significant reduction (p <0.05) in the egg protein content was observed in baked biscuits. With 25 minutes of baking, were evidenced reductions of 83.1; 92.6 and 100% for the temperatures 150, 180 and 210 ° C, respectively, indicating influence of the processing on the antigenic potential. The results of the quantitative approach of validation showed that the performance criteria were not complied. In the qualitative approach, 100 % of sensitivity and reliability rates were found for the blank samples. In the levels 0.125; 0.185 and 0.25 mg/kg the sensitivity and reliability rates were 0; 90 and 80 %, respectively. From the level 0.5 mg/kg, the estimated values of these rates were 100 %, demonstrating sensitivity of the method. The accordance ranged from 0.5 to 1.0 and concordance from 0.7 to 1.0. For the concentrations of 0; 0.125 and over 0.5 mg/kg the concordance reaches maximum values, showing standardization of the method. The detection limit was established as 0.2 mg/kg. The studied kit presented selectivity in the presence of the other allergenic protein beta-lactoglobulin. Thus, the baking conditions were evidenced as critical for the antigenicity of egg white proteins and the studied kit was considered appropriate for the detection of egg white proteins in semi-sweet biscuits, characterizing an important tool for the implementation of the control of food allergens labeling.
publishDate 2017
dc.date.none.fl_str_mv 2017-02-23
2019-08-10T22:22:14Z
2019-08-10T22:22:14Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
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dc.identifier.uri.fl_str_mv http://hdl.handle.net/1843/BUOS-BATFN3
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dc.language.iso.fl_str_mv por
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dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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dc.publisher.none.fl_str_mv Universidade Federal de Minas Gerais
UFMG
publisher.none.fl_str_mv Universidade Federal de Minas Gerais
UFMG
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFMG
instname:Universidade Federal de Minas Gerais (UFMG)
instacron:UFMG
instname_str Universidade Federal de Minas Gerais (UFMG)
instacron_str UFMG
institution UFMG
reponame_str Repositório Institucional da UFMG
collection Repositório Institucional da UFMG
repository.name.fl_str_mv Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)
repository.mail.fl_str_mv repositorio@ufmg.br
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