Screening of stereoisomeric chloramphenicol residues in honey by ELISA and CHARM ® II test – the potential risk of systematically false-compliant (false negative) results

ABSTRACT Chloramphenicol (CAP) is a broad-spectrum antibiotic used widely both in human and in veterinary medication but due to adverse health effects is not authorised anymore for use in food-producing animals in many countries. CAP molecule contains two asymmetric centers resulting in four para-CAP stereoisomers, but only the RR-CAP enantiomer is bioactive with significant antimicrobial activity. In this study the detection of the four CAP stereoisomers was tested by five commercial ELISA kits and the Charm® II Chloramphenicol Test. These immunoassay tests are commonly used and widely accepted for screening of CAP residues in foods of animal origin, including honey. The test results definitely show that SS-CAP residues are not detected; even high SS-CAP concentrations are missed due to the lack of any cross reactivity and the high specificity of the CAP antibodies to RR-CAP. In former studies chiral LC-MS/MS analysis indicated clearly that honey samples with raised CAP concentrations often contain the SS-CAP enantiomer in addition to the bioactive RR-CAP. According to this study, the investigated screening tests carry the risk of systematically false-compliant (false negative) results for CAP and a discrepancy between LC-MS/MS and ELISA/Charm® test results. As a consequence of this study, it is recommended that immunoassay manufacturers develop and use CAP antibodies which also bind SS-CAP. The origin of SS-CAP residues in honey samples is discussed and general toxicological and regulatory aspects of CAP stereoisomers are raised. Graphical abstract