An analytical validation of enzyme-linked immunosorbent assay (ELISA) screening for detection of chloramphenicol (CAP) in shrimp and fish was conducted according to the Commission Decision 2002/657/EC and guidelines for the validation of screening methods for residues of veterinary medicines. The analyte was extracted from shrimp and fish with ethyl acetate mixture, and CAP concentrations were measured photometrically at 450 nm. The recovery rate of the analyte from spiked samples was 80%. For the laboratory the cut-off level of CAP in fish and shrimp as the minimum recovery was established along with detection capability (CCβ). No relevant interferences between matrix effects and structurally related substances including florfenicol and thiamphenicol were observed. The experimental results were quite satisfactory and ELISA method was found very useful for determination of CAP residues in shrimp and fish monitoring.
A simultaneous and fast analytical method was developed for the identification of fournitrofuran metabolites from the fish and shrimp samples. Homogenized samples were hydrolyzedand derivatized with 4-nitrobenzaldehyde. Subsequently, extracted with ethyl acetate, evaporatedto dryness and the residue was re-dissolved in Hexane. Commercial enzyme-linkedimmunosorbent assay (ELISA) method was applied for the analysis of nitrofuran metabolites. Themethod was validated in shrimp and fish matrix according to the criteria defined in CommissionDecision 2002/657/EC for qualitative screening method following the guidelines set by thecommunity reference laboratories residues (CRLs) 2010. Characteristic’s parameters as detectioncapability (CC?), specificity/selectivity, stability, recovery and precision were determined.Detection capability (CC?) for nitrofuran metabolites (AMOZ, AOZ, AHD and SEM) in Fishand shrimp matrix was in the range of 0.5- 0.75?g/kg, which were less than the MinimumRequired Performance Limit (MRPL) of 1?g/kg set by European Union. The proposed method issuitable for semi-quantitative screening analysis of antibiotics in the fish and shrimp muscle inconformity with the current EU performance requirements before exporting to EU and othercountries. Results from analysis of unknown samples by the developed ELISA method werecomparable to those obtained by a liquid chromatography-tandem mass spectrometry (LCMS/MS) method. Accuracy and precision of the method had also been checked through participation of International Proficiency Testing (PT) having a very satisfactory performance score.
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