Several studies have demonstrated that fish feeds contain significant concentrations of chemical contaminants, many of which can bioaccumulate and bioconcentrate in fish tissues (1). The serious concern regarding the use of fish meal and fish oil in the the aquaculture industry has led to extensive search of alternative raw materials for aquafeeds. The most obvious alternatives are oils and proteins of plants origin. The use of these alternative feed ingredients can introduce contaminants that were previously not associated with fish farming such as mycotoxins (2). Ochratoxin A (OTA) is a mycotoxin produced as a secondary metabolite by various Aspergillus and Penicillium species with nephrotoxic, carcinogenic, immunotoxic and teratogenic potential (3). OTA has been found in several food commodities, including cereals and can also be present in food of animal origin as a result of carryover from contaminated feed (3). The aim of the present study was to determine OTA concentrations in muscle, kidney and liver of 10 seabream and 10 seabass of farmed origin collected on the market. Analysis will be performed by using an enzymatic digestion (ED) method coupled to high-performance liquid chromatography with a fluorescence detector (HPLC-FLD). Fish tissues were digested for 1 hour at 37°C with a 1% pancreatin solution in a phosphate buffer and then cleaned up with ethylacetate. After being evaporated to dryness and re-dissolved, the sample was processed using HPLC-FLD. The method was validated for: specificity, recovery, trueness, selectivity, linearity, limit of detection (LOD) and limit of quntification (LOQ), repeatability and reproducibility. Recoveries of analytical method were higher than 85 % for all the matrices. Intra and inter-day repeatability expressed as relative standard deviation were less than 9%. The LOD and LOQ for liver and muscles samples were 0.001 and 0.002 μg/kg, respectively. The LOD and LOQ for kidney samples were 0.01 and 0.02 μg/kg, respectively. The highest concentrations of OTA were found in kidney of the 20 fishes analyzed (range
DETERMINATION OF OCHRATOXIN A IN FARMED FISH BY ENZYMATIC DIGESTION (ED) COUPLED TO HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH A FLUORESCENCE DETECTOR (HPLC-FLD)
Meucci Valentina;Luci Giacomo;Armani Andrea;Intorre Luigi
2018-01-01
Abstract
Several studies have demonstrated that fish feeds contain significant concentrations of chemical contaminants, many of which can bioaccumulate and bioconcentrate in fish tissues (1). The serious concern regarding the use of fish meal and fish oil in the the aquaculture industry has led to extensive search of alternative raw materials for aquafeeds. The most obvious alternatives are oils and proteins of plants origin. The use of these alternative feed ingredients can introduce contaminants that were previously not associated with fish farming such as mycotoxins (2). Ochratoxin A (OTA) is a mycotoxin produced as a secondary metabolite by various Aspergillus and Penicillium species with nephrotoxic, carcinogenic, immunotoxic and teratogenic potential (3). OTA has been found in several food commodities, including cereals and can also be present in food of animal origin as a result of carryover from contaminated feed (3). The aim of the present study was to determine OTA concentrations in muscle, kidney and liver of 10 seabream and 10 seabass of farmed origin collected on the market. Analysis will be performed by using an enzymatic digestion (ED) method coupled to high-performance liquid chromatography with a fluorescence detector (HPLC-FLD). Fish tissues were digested for 1 hour at 37°C with a 1% pancreatin solution in a phosphate buffer and then cleaned up with ethylacetate. After being evaporated to dryness and re-dissolved, the sample was processed using HPLC-FLD. The method was validated for: specificity, recovery, trueness, selectivity, linearity, limit of detection (LOD) and limit of quntification (LOQ), repeatability and reproducibility. Recoveries of analytical method were higher than 85 % for all the matrices. Intra and inter-day repeatability expressed as relative standard deviation were less than 9%. The LOD and LOQ for liver and muscles samples were 0.001 and 0.002 μg/kg, respectively. The LOD and LOQ for kidney samples were 0.01 and 0.02 μg/kg, respectively. The highest concentrations of OTA were found in kidney of the 20 fishes analyzed (rangeFile | Dimensione | Formato | |
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