Abstract

A method was developed and validated in-house for determination of ochratoxin A (OTA) in wine using charge coupled device (CCD) to acquire the fluorescence images of mycotoxin from high-performance thin-layer chromatography (HPTLC) under UV lamp. The method showed a mean recovery of 90.4%. The quantification and detection limist were 0.1 µg L-1 and 0.016 µg L-1 per spot, respectively. The results of validation confirmed the efficiency of the method, which is sensitive enough to be used to quantify OTA in wine. The occurrence of OTA in Brazilian wines was evaluated. OTA was found in one sample at a level of 4.5 µg L-1, which is higher than the limit of 2 µg L-1 considered acceptable by the Scientific Commission of the European Communities. This study demonstrated the applicability of HPTLC using CCD as a tool to determine OTA in wine.

Highlights

  • Ochratoxin A (OTA), 7-(L-b-phenylalanyl-carbonyl)carboxyl-5-chloro-8-hydroxy-3,4-dihydro-3R-methyl­ isocumarin, is a mycotoxin produced by Penicillium verrucosum, Aspergillus ochraceus,[1] A. niger and A. carbonarius.[2,3] OTA occurs in several food products such as in cereals,[4] coffee beans,[5] beer,[6] cacau,[7] wheat,[8] dried fruit,[9] cheese,[10] grape juice and wine.[11]

  • The sensitivity of the high-performance thin-layer chromatography (HPTLC)-charge coupled device (CCD) technique was evaluated by determining limit of detection (LOD) for OTA, which was derived from the fluorescence of OTA standard with the lowest detectable signal

  • This technique is based in use of CCD to acquire the fluorescence images of OTA from HPTLC plate under UV lamp

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Summary

Introduction

Ochratoxin A (OTA), 7-(L-b-phenylalanyl-carbonyl)carboxyl-5-chloro-8-hydroxy-3,4-dihydro-3R-methyl­ isocumarin, is a mycotoxin produced by Penicillium verrucosum, Aspergillus ochraceus,[1] A. niger and A. carbonarius.[2,3] OTA occurs in several food products such as in cereals,[4] coffee beans,[5] beer,[6] cacau,[7] wheat,[8] dried fruit,[9] cheese,[10] grape juice and wine.[11]. OTA is receiving major attention for its nephrotoxic effects. This toxin is known for its mutagenic,[13] teratogenic,[14] immunosuppressive[15] and carcinogenic proprieties.[16,17,18,19] OTA has been associated with Balkan. There are recommendations of Provisional Tolerable Weekly Intakes (PTWI) for OTA of 100 ng kg-1 body weight.[1] The Scientific Commission of the European Community established the maximum allowed level of 2 μg L-1 of OTA in wine.[23] There is yet no maximum permissible level established for this mycotoxin in wine in Brazil, but it is necessary to adopt limits for OTA so as to minimize the health hazard risk. After the first detection of OTA in wine,[24] several surveys were conducted in several countries to examine the true occurrence of this toxin in wine[25,26,27] as well

Determination of Ochratoxin A in Wine
Ochratoxin A standard solution
Ochratoxin A extraction
CCD Imaging system
Method validation
Results and Discussion
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