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Ciência e Técnica Vitivinícola

versão impressa ISSN 0254-0223

Ciência Téc. Vitiv. v.23 n.1 Dois Portos  2008

 

Validation study of immunochemical elisa assay for ochratoxin a quantification in dessert wines from sun-dried grapes

 

F. J. E. Alcaide1, S. A. Aguilar1

1 Quality Department. Navisa Industrial Vinícola Española S.A. Ctra. de Montalbán s/n. -14550 Montilla, Córdoba, Spain. E-mail.: calidad@navisa.es

(Manuscrito recebido em 24.03.08. Aceite para publicação em 11.06.08)

 

 

ABSTRACT

Ochratoxin A (OTA) contents have implications on consumers safety and wine quality. Immunological methods are widely used to determine OTA in various matrices. Solid-phase extraction with immunoaffinity columns containing antibodies specific to ochratoxin A were used to clean-up the samples by two different methods in alkaline medium. Competitive enzyme-linked immunosorbent assay (ELISA) commercial kit was utilized for OTA quantification in dessert wines samples. This ELISA assay uses an instrumental response based on absorbance measurements at 450 nm. The method was performed with standard solutions (provided with the kit), in spiked and naturally contaminated wine samples. The detection limit was 0.054 ìg/L and recoveries of OTA from spiked wine samples, at levels from 1 to 3 ìg/L, ranged from 94 to 102%, with relative standard deviations less than 3%. This ELISA method was checked against an official instrumental method like HPLC with tandem mass spectrometric and with fluorescence detection. ELISA kit method resulted effective for measuring OTA ranging from 0.25 to 9 ìg/L in dessert wines.

Keywords: Ochratoxin A; ELISA; Wine; Mycotoxins; Immunochemical method.

 

 

RESUMO

Estudo da validação do método imunoquímico ELISA para a quantificação deocratoxina A em vinhos doces de sobremesa elaborados com usas desidratadas ao sol

Os conteúdos do ocratoxina A (OTA) têm implicações para as qualidades dos vinhos, revelando-se um parâmetro importante para a segurança alimentar e consecuentes implicações para a saúde dos consumidores. Os métodos imunológicos são amplamente utilizados para determinação da ocratoxina A em várias matrizes. A extracção fase-sólida com colunas de imunoafinidade contendo anticorpos específicos para ocratoxina A foram utilizados para a limpeza de amostras por dois métodos diferentes, em meio alcalino. O kit comercial ELISA utilizado na quantificação de OTA em amostras do vinhos doces mostrou-se adequado. Este ensaio ELISA utiliza uma resposta instrumental baseada em medições do absorvência a 450 nm. Foram usadas as soluções padrão provenientes do kit, amostras de vinho doce reforçado e amostras naturalmente contaminadas. O limite de detecção foi de 0.054 ìg/L e as taxas de recuperação de OTA de amostras vinho reforçado de 1 a 3 ìg/L, com variações entre 94 e 102%, e desvios padrão relativos inferiores a 3%. O método ELISA foi comparado com o método cromatografia líquida de alto desempenho e detecção por espectrómetro de massa (MS/MS) e com detector de fluorescência. O kit ELISA revelou-se o método mais eficaz para medir OTA numa gama de 0.25 a 9 ìg/L em os vinhos doces.

Palavras Chave: Ocratoxina A; ELISA; Vinho; Micotoxinas; Método imunoquímico.

 

 

Texto completo disponível apenas em PDF.

Full text only available in PDF format.

 

 

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