Analysis of chloramphenicol in honeys of different geographical origin by liquid chromatography coupled to electrospray ionization tandem mass spectrometry

Abstract

A sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed to detect trace amounts of the antibiotic chloramphenicol (CAP) in honey. The methodology entailed a solid-phase extraction of aqueous honey solutions followed by liquid-liquid partitioning, filtration and direct injection onto the LC-MS/MS system. Honey extracts were spiked with an isotopically labelled internal standard (d(5)-CAP) to compensate for analyte loss and potential ion suppression during the MS stage. Detection of the analyte was achieved by negative ionization electrospray in the selected reaction monitoring (SAM) mode. For confirmation, four characteristic mass transitions were monitored each for the analyte and the surrogate standard. The method was validated according to the latest European Union criteria for the analyses of veterinary drug residues in food. At all three fortification levels studied (0.1, 0.2, 0.5 microg kg(-1)) the method was accurate to within 15%. The repeatability and within-laboratory reproducibilities were <12 and 18%, respectively. The decision limit (CC alpha) and detection capability (CC beta) were both <0.1 microg kg(-1). The procedure provides a sensitive and reliable method for the determination of residues of chloramphenicol in honey. Numerous raw honeys of various geographical origins were analysed, showing extensive contamination particularly those of Chinese origin.

Contaminants of bee products

“…Results from different laboratories showed that a great part of Chinese honey, but also of honey from various countries, contains chloramphenicol in quantities greater than the EU MRL of 0.3 µg/kg (e.g. Reybroeck, 2003;Verzegnassi et al, 2003). Recently, nitrofuranes were also found in honey (Stiftung Warentest, 2004).…”

“…Sulfonamides sulfathiazole, sulfamerazine, sulfamethazine, sulfamethaxazole, sulfadiazine, sulfamethoxypiridazine, sulfadoxine, sulfadimidine, sulfanilamide (Martel and Zeggane, 2003;Reybroeck, 2003;Wallner, 2003;Kaufmann and Känzig, 2004) Aminoglycosides streptomycine, dihydrostreptomycine (Morlot and Beaune, 2003;Reybroeck, 2003;van Bruijnsvoort et al, 2004) Tetacyclines tetracycline, oxytetracycline, chlortetracycline, doxycycline (Argauer and Moats, 1991;Tantillo et al, 2000;Morlot and Beaune, 2003;Reybroeck, 2003;Sabatini et al, 2003) Amphenicols chloramphenicol (Dharmananda, 2003;Reybroeck, 2003;Verzegnassi et al, 2003;Ortelli et al, 2004) Macrolides tylosine myrosamine (Baggio et al, 2004;Feldlaufer et al, 2004) (Nakajima et al, 1998) Beta-lactams penicillins (Nakajima et al, 1997;Reybroeck, 2003) Nitrofuran metabolites AOZ, SC (Stiftung Warentest, 2004;Jenkins and Young, 2005) AOZ: 3-amino-2-oxazolidinone; SC: semicarbazide.…”

Study of the distribution and depletion of chloramphenicol residues in bee products extracted from treated honeybee (Apis melliferaL.) colonies

“…CAP is listed as a prohibited substance for use in food producing animals with an Annex IV classification under European legislation (Council Regulation 2377/90) There is evidence that CAP is effective in controlling Paenibacillus larvae larvae which can cause American Foulbrood Disease (AFB) (Kochansky et al, 2001) and it has been used for this purpose. In a recent survey 97% of samples of honey from China contained residues of CAP (Verzegnassi et al, 2003). Similarly a high incidence of CAP residues has been reported in royal jelly (Calvarese et al, 2006;Ishh et al, 2006).…”

Scientific Opinion on Chloramphenicol in food and feed

“…None of the Australian honeys contained chloramphenicol, while 97 % of the Chinese samples contained chloramphenicol with an average chloramphenicol concentration of 4.8 µg/kg (range: 0.1-75 µg/kg). Intermediate results were observed for the samples from Argentina (8 % positive samples, mean concentration 0.1 µg/kg), Cuba (6.3 % positive samples, mean concentration 0.3 µg/kg) and Thailand (15 % positive samples, mean concentration 1.4 µg/kg) (Verzegnassi et al, 2003).…”