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http://hdl.handle.net/1843/URMR-7RYPJ5| Type: | Dissertação de Mestrado |
| Title: | Otimização e validação de método para determinação de histamina em pescado |
| Authors: | Tarliane Maria da Silva |
| First Advisor: | Maria Beatriz Abreu Gloria |
| First Referee: | David Lee Nelson |
| Second Referee: | Edgar de Alencar Teixeira |
| Third Referee: | Luis Antonio Minim |
| Abstract: | A exportacao de pescado fresco para a Europa tem sido limitada pela exigencia de analises laboratoriais de histamina. As analises devem ser realizadas por cromatografia liquida de alta eficiencia (CLAE). O metodo sugerido, entretanto, faz uso de acido perclorico, extrator com potencial explosivo, depende de reacao de derivacao da histamina, a qual exige manipulacao morosa e excessiva da amostra, aumentando a possibilidade de perdas em recuperacao. Alem disso, pelo fato da analise ser demorada, corre-se o risco de prejudicar a exportacao do material fresco, cuja vida de prateleira e limitada. Visando atender estas necessidades, teve-se com este trabalho o objetivo de otimizar e validar um metodo eficiente e rapido por CLAE para determinacao de histamina em pescado. A extracao de histamina foi otimizada, definindo-se como 70 s o tempo de agitacao; 11250 x g, 3 min e 0 C a velocidade, otempo e a temperatura de centrifugacao, respectivamente; e o tempo de extracao foi reduzido de 90 para 15 minutos, em relacao a extracao originalmente utilizada. O padrao interno 1,3-diaminopropano recomendado por DUFLOS et al. (1999), co-eluiu com a feniletilamina e putrescina, sendo sua utilizacao inviavel. Comparando-se os metodos de DUFLOS et al. (1999) e adaptado de VALE & GLORIA (1997), observou-se que o ultimo e o mais apropriado a analise de histamina em pescado, sendo, portanto, escolhido para otimizacao e validacao. O metodo validado para determinacao de histamina em pescado mostrou-se linear na faixa de trabalho de 0,2 a 60 Êg/mL, semefeito da matriz, seletivo, exato e preciso. Em virtude dos limites estabelecidos pela Uniao Europeia se enquadrarem na faixa de 100 a 200 mg/kg, o metodo e viavel para analise de histamina em pescado, apresentando reducao do tempo total de analise em relacao aos dois metodos comparados. A partir do metodo otimizado e validado foramavaliados os teores de histamina em amostras de atum fresco e enlatado comercializadas em Belo Horizonte, verificando-se que estes estao de acordo com oslimites regulamentados pelo Mercosul, FDA e Uniao Europeia. |
| Abstract: | The exportation of fresh fish to Europe has been limited by therequirement of certificates with the levels of histamine. The method recommended for the analysis is high performance liquid hromatography (HPLC). Histamine is extracted with perchloric acid and derivatized prior to analysis. However, the extractor is explosive and the derivatization reaction requires excessive handling which can affectnegatively the efficiency of the method. Furthermore, since the analysis takes a long time, the exportation of fresh fish is limited. In order to solve such problems, the objective of this work was to optimize and validate an efficient and rapid HPLC method for the determination of histamine in fish. The extraction was optimized and the defined parameters were sample homogenization for 70 s; 11250 x g, 3 min and 0 C for thespeed, time and temperature of centrifugation, respectively. Under these conditions, the extraction time was reduced from 90 to 15 minutes, compared to the recommended method. The internal standard (1,3-diaminopropane) recomended by DUFLOS et al. (1999), co-eluted with phenylethylamine and putrescine, and, therefore, could not beused. Comparing the analytical procedures used by DUFLOS et al. (1999) and adapted from VALE & GLORIA (1997), the last one was the most appropriate for the analysis of histamine in fish, and it was chosen for further studies. The method was optimized and validated: it was linear in the range of 0.2 to 60 Êg/mL; there was no effect of the matrix;and the method was selective, exact and precise. The method was considered viable for the analysis of histamine in fish as recommended by the Europeian Union which requires limits of 100 to 200 mg/kg. Furthermore, it could be performed in a shorter period of time compared to the original method. Using this method, the levels of histamine were determined in fresh and canned tuna samples commercialized in /beloHorizonte, state of Minas Gerais, Brazil. Every sample analyzed was in compliance with the Mercosur, FDA and Europeian Union standards. |
| Subject: | Cromatografia líquida de alta eficiência Alimentos Contaminação Pescados Análise Pescados Contaminação Histamina |
| language: | Português |
| Publisher: | Universidade Federal de Minas Gerais |
| Publisher Initials: | UFMG |
| Rights: | Acesso Aberto |
| URI: | http://hdl.handle.net/1843/URMR-7RYPJ5 |
| Issue Date: | 7-Oct-2008 |
| Appears in Collections: | Dissertações de Mestrado |
Files in This Item:
| File | Description | Size | Format | |
|---|---|---|---|---|
| disserta__o_tarliane.pdf | 3.74 MB | Adobe PDF | View/Open |
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