Abstract
A rapid fluorescence polarization immunoassay (FPIA) was optimized and validated for the determination of ochratoxin A (OTA) in rye and rye crispbread. Samples were extracted with a mixture of acetonitrile/water (60:40, v/v) and purified by SPE-aminopropyl column clean-up before performing the FPIA. Overall mean recoveries were 86 and 95% for spiked rye and rye crispbread with relative standard deviations lower than 6%. Limits of detection (LOD) of the optimized FPIA was 0.6 μg/kg for rye and rye crispbread, respectively. Good correlations (r > 0.977) were observed between OTA contents in contaminated samples obtained by FPIA and high-performance liquid chromatography (HPLC) with immunoaffinity cleanup used as reference method. Furthermore, single laboratory validation and small-scale collaborative trials were carried out for the determination of OTA in rye according to Regulation 519/2014/EU laying down procedures for the validation of screening methods. The precision profile of the method, cut-off level and rate of false suspect results confirm the satisfactory analytical performances of assay as a screening method. These findings show that the optimized FPIA is suitable for high-throughput screening, and permits reliable quantitative determination of OTA in rye and rye crispbread at levels that fall below the EU regulatory limits.
Highlights
Several Penicillium and Aspergillus species growing in different agricultural commodities in the field or during storage can produce ochratoxin A (OTA) [1], a mycotoxin that has recently received a special focus due to its toxic effects
The evaluation of the matrix effect on the fluorescence polarization immunoassay (FPIA) was performed by using spiked extracts of rye and
These results showed that the limit of detection (LOD) obtained was far below the maximum permitted levels of quantification rye and rye cereals crispbread
Summary
Several Penicillium and Aspergillus species growing in different agricultural commodities in the field or during storage can produce ochratoxin A (OTA) [1], a mycotoxin that has recently received a special focus due to its toxic effects. OTA has multiple toxic effects that are a real menace to the health of humans and animals. Many studies have indicated that this mycotoxin shows nephrotoxic, hepatotoxic, embryotoxic, teratogenic, neurotoxic, immunotoxic, genotoxic, and carcinogenic effects on several species [2]. OTA has been extensively reported to contaminate a wide variety of foods and beverages such as cereals and derived products, coffee, spices, grapes, beer and wine at global level [5,6,7,8]. According to recent reports on food incidence of OTA, cereals and their cereal-based products contribute to Toxins 2017, 9, 305; doi:10.3390/toxins9100305 www.mdpi.com/journal/toxins
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