Desenvolvimento e validação de metodologia analítica para a determinação de micotoxinas em vinhos

Abstract

Mycotoxins are secondary metabolites produced by filamentous fungi that occur naturally in foods. The mycotoxins most commonly found in fruits and their processed products are alternariol (AOH), alternariol monometyl ether (AME), citrinin (CIT), ochratoxin (OTA) and patulin (PAT). OTA is produced by Penicillium and Aspergillus species and have nephrotoxic, mutagenic, teratogenic and carcinogenic. Often, OTA is present in foods in co-occurrence with other mycotoxin, the citrinin, which also have nephrotoxic and teratogenic. PAT is mainly produced by Penicillium species and occurs in a wide variety of fruits such as apple, grape and plum. AOH and AME are produced by Alternaria alternata and have genotoxic and carcinogenic properties. Along with OTA, mycotoxins are more common in wines. Given the importance of surveillance of mycotoxins in foods, as soon as regulations established by ANVISA, has developed two analytical methods for detection of five mycotoxins that may be present in grapes and consequently in wine. The first method was developed and validated for analysis of patulin using high performance liquid chromatography coupled tandem mass spectrometry (HPLC-MS/MS) with atmospheric pressure chemical ionization (APCI). The sample preparation prior to analysis, it moved by extraction with ethyl acetate and vortex, freezing the extract for 24 hours, solvent evaporation in a water bath, resuspended and diluted. The limit of quantitation (LQ) obtained was 12 μg kg-1 and the average recovery of the method was 89.4%. Through this methodology, we analyzed 111 samples of wines and any sample no showed patulin contamination. The second method was developed and validated for simultaneous analysis of AOH, AME, CIT and OTA using HPLC-MS/MS with electrospray ionization (ESI). For preparing for analysis, the sample was diluted with solution of methanol and water. The LQ obtained was 1 μg kg-1 for AOH, AME and OTA and 20 μg kg-1 for CIT. The average recovery of the method was 92.5% for AOH, 96.1% for AME, 97.8% for OTA and 96.2% for CIT. By analyzing 111 samples of wine, found 81 contaminated with AOH, 48 with AME, 2 with OTA and no with CIT.