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  • Title: Alternatives for sample pre-treatment and HPLC determination of Ochratoxin A in red wine using fluorescence detection.
    Author: Tessini C, Mardones C, von Baer D, Vega M, Herlitz E, Saelzer R, Silva J, Torres O.
    Journal: Anal Chim Acta; 2010 Feb 15; 660(1-2):119-26. PubMed ID: 20103152.
    Abstract:
    Ochratoxin A is a mycotoxin widely studied due to its nephrotoxic, immunotoxic, teratogenic and carcinogenic effects. The European Commission has fixed maximum limits for Ochratoxin A in wines and in other foods. In order to determine Ochratoxin A levels in red wine, the present paper contrasts and discusses the results of a systematic study of analytical parameters for sample pre-treatment using different immunoaffinity cartridges as well as C-18 cartridges with three solvent combinations. The direct injection of wine into two types of C-18 chromatographic columns (conventional packed column and monolithic column) is evaluated as screening method. In all cases, the analysis was carried out using HPLC with fluorescence detection. The results show statistical differences when 3 types of immunoaffinity columns were used, while higher recoveries were obtained for C-18 cartridges using acetonitrile as extraction solvent. Repeatability and accuracy of immunoaffinity and C-18 sample pre-treatment were statistically comparable (alpha=0.05). Their sensitivity was also comparable, although more favorable detection limits were obtained using the immunoaffinity treatment (0.01 microg L(-1)) in comparison with C-18 treatment (0.09 microg L(-1)). Considering the maximal allowed concentration of Ochratoxin A in wine (2.00 microg L(-1)), both methods are suitable for its determination in wine. Both methods were applied to determine this toxin in 154 wine samples, and the quantitative results demonstrated statistic comparability (alpha=0.05). These results were also confirmed from the qualitative point of view using a GC-MS method. To find an easy screening method, based on a recent publication, a monolithic HPLC column and 2 conventional packed columns were tested for Ochratoxin A determination in real wine samples by direct injection, without previous clean-up. The results show that this procedure is not useful at the concentration levels usually found in wine and although shorter time is required when using the monolithic columns even with the chromatographic analysis. Finally, based on the results, it was concluded that the combination of C-18 cartridges with conventional particle packed columns and HPLC-FLD is the most appropriate alternative for Ochratoxin A analysis in wine. Indeed, considering cost, sensitivity and selectivity, this method can be used in broad prospective programs.
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