ELISA qualitative screening of chloramphenicol in muscle, eggs, honey and milk: method validation according to the Commission Decision 2002/657/EC criteria

Abstract

A commercial enzyme-linked immunosorbent assay (ELISA) test has been applied for the qualitative screening analysis of chloramphenicol (CAP) in food of animal origin (muscle, eggs, milk and honey) at level corresponding to the European Union (EU) minimum required performance limit (MRPL) set for CAP analysis. Different analytical protocols were developed for the extraction and purification of CAP from the matrices under investigation. The ELISA kit validation study was performed according to the Commission Decision 2002/657/EC criteria established for qualitative screening methods. In this regard, the following parameters were determined: detection capability (CCβ), specificity/selectivity, applicability/ruggedness/stability. The CCβ value resulted below the MRPL (0.3 μg kg −1) for all the examined matrices. No relevant interferences from matrix effects or structurally related substances (florfenicol (FF) and thiamphenicol (THP)) were observed. Slight variations of some critical factors in the sample pre-treatment and clean up for muscle were deliberately introduced for ruggedness evaluation and they did not result in any negative effect on the CAP detection. In addition, the recovery rates and precision at three concentration levels (0.30–0.45–0.60 μg kg −1) were evaluated for all matrices under investigation. The proposed method is suitable for qualitative screening analysis of CAP in the above-mentioned food in conformity with the current EU performance requirements.