Simultaneous and fast determination of antibiotics as nitrofuran metabolites in fish and shrimp muscle using Enzyme-Linked Immunosorbent Assay (ELISA)

Abstract

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.