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Ochratoxin A: Comparison of Extraction Methods from Grapes and Quantitative Determination by Different Competitive Enzyme-Linked Immunosorbent Assay Kits

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Abstract:

The European Community has recently established a maximum limit for ochratoxin A (OTA) concentration in grapevine products, but many practical difficulties remain concerning the establishment of optimum cost-effective methods of quantification. The performance of four extraction procedures and three commercial competitive enzyme-linked immunosorbent assays (cELISAs) for grapes were compared. Results differed for the extractions and the cELISA kits. The advantage of using immunoaffinity columns (IACs) in the extraction was the excellent detection limit, which was between 0.06 and 0.0075 ng ml−1 depending on the cELISA kit used. Despite lower sensitivity (between 1.2 and 0.15 ng ml−1 depending on the cELISA kit), an extraction method in liquid phase, which was simple and inexpensive, was confirmed as suitable for quantifying OTA at levels estimated to be dangerous for human health. Two of the three cELISA kits produced satisfactory results. When these two cELISAs were coupled with IAC extraction, the lower quantification limits were 0.010 and 0.0075 ng ml−1, respectively, and the dynamic ranges were 50 and 27, respectively. The most reliable procedures were then compared with the reference method, high-performance liquid chromatography plus fluorescent detection coupled with an IAC. The results were very similar, although the cELISAs generally provided slightly higher values than did the chromatography method. The IAC method coupled with the cELISA was four times more sensitive than was the IAC method coupled with the chromatography method. The cELISA detection techniques were excellent alternatives to the already established chromatographic protocols, especially for mass screening and for determining concentrations of OTA as low as 0.010 ng ml−1.

Document Type: Research Article

Affiliations: 1: C.R.A.-VIT Centro per la Ricerca in Viticoltura, Viale XXVIII Aprile 26, 31015 Conegliano (TV), Italy 2: C.R.A.-UTV Unità di Ricerca per l'uva da tavola e la vitivinicoltura in ambiente mediterraneo, Via Vittorio Veneto 26, 70051 Barletta (BA), Italy

Publication date: December 1, 2008

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