Determination of lincomycin and tylosin residues in honey by liquid chromatography/tandem mass spectrometry

Abstract

A simple and rapid analytical method was developed for the determination of lincomycin and tylosin residues in honey as part of field studies examining the efficacy and target animal safety of these antibiotics to control American foulbrood disease in honey bees. Residues of the antibiotics were determined using liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS). Honey samples were diluted and injected directly into the LC/MS/MS system without additional cleanup by solid-phase extraction or liquid-liquid partitioning. A six-port valve system was utilized to selectively route eluant from the LC column into the mass spectrometer only during a relatively short portion of the chromatographic run corresponding to the elution of the analytes of interest. Minimal contamination of the MS source chamber was observed despite the analysis of large numbers of samples. Using internal standard quantitation, excellent accuracy and precision were obtained with no apparent matrix-to-matrix variation. Based on the analysis of fortified replicates, the mean percent deviation from the theoretical concentration and the percent relative standard deviation were both less than 10% for tylosin over an analytical range of 10-1000 microg/kg. Slightly higher mean percent deviations and relative standard deviations were observed for the analysis of lincomycin in fortified replicate samples. The method detection limits were determined to be 5 and 2 microg/kg for lincomycin and tylosin, respectively.

Aspectos analíticos e regulatórios na determinação de resíduos de macrolídeos em alimentos de origem animal por cromatografia líquida associada à espectrometria de massas

“…Um solvente mais apolar (acetato de etila) foi adicionado ao final do gradiente para remover da coluna resíduos de mel que não seriam eluídos com acetonitrila. 36,37 Em outro caso, macrolídeos e outros antimicrobianos, num total de 19 substâncias, foram extraídos de músculo e fígado de suínos, bovinos e equinos com metanol a 70%, com posterior diluição em cinco vezes com água e injeção no sistema LC-MS. Este método foi adequado para monitorar os antimicrobianos nestas matrizes e conseguiu detectar macrolídeos com relação sinal/ ruído maior que 8 em concentrações de 50 a 1000 mg kg -1 . 30 Um terceiro exemplo envolve a análise de eritromicina em trutas, onde 1 g de amostra foi misturada com água (1 mL) e o analito foi extraído a partir da adição de 4 mL de acetonitrila, agitação, sonicação e centrifugação a 10000 g. O sobrenadante foi coletado e o pelete (precipitado) foi ressuspendido em 5 mL de acetonitrila e agitado, sonicado e centrifugado novamente e o sobrenadante coletado.…”

Current trends in sample preparation for growth promoter and veterinary drug residue analysis

“…Thompson et al investigated the fate of tylosin residues in the honey from treated bees [294]. This study showed that tylosin A degrades to yield the antimicrobially active degradation product tylosin B, also known as desmycosin.…”

Trace analysis of antibacterial tylosin A, B, C and D in honey by liquid chromatography-electrospray ionization-mass spectrometry

“…To solve this problem, different mobile phases, suggested in the bibliography for the analysis of TA and other macrolides, were tested. There are mainly ternary mobile phases consisting of mixtures of ACN-formic acid-water [10]; methanol-01% v/v TFA in ACN or 0.1% v/v TFA in water [22]; methanol-ACN-oxalic acid [24]; ACN-heptafluorobutiric acid-ethyl acetate [23]; water-potassium phosphate buffer-THF [26]. Also, some binary mobile phases have been proposed [21,27].…”

“…To suppress these interferences, a washing step was necessary. Taking into account different modes to solve this problem [10,22,24], some water-methanol mixtures (with 20, 40, 60, 80 and 100% of methanol) were assayed on fortified honey. The interferences were eliminated with F40% of methanol.…”

“…With regard to the sample preparation, the most common procedures include a dilution of sample at alkaline pH and an isolation step based on the use of SPE on C18 or polymeric cartridges [10,[21][22][23]. This step could be omitted if MS/MS was employed, as it has been recently proposed [24] for TA, although unfortunately this is not a common instrumentation in control laboratories.…”