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High sensitivity quantitation of metabolites of nitrofuran antibiotics in animal tissue using LC/MS/MS

A sensitive and simple high performance liquid chromatograph tandem mass spectrometry (LC/MS/MS) method is presented for a simultaneous analysis of the metabolites of four nitrofuran antibiotics in chicken tissue.

Abstract

The animal meat samples were prepared by homogenization, organic solvent washes, hydrolysis and derivatization followed by liquid-liquid (L-L) extraction before analysis. The limits of detection were 0.05-0.15 ng/g animal tissue using a triple-quad mass spectrometer with electrospray ionization in a positive mode. The analyte quantitation and identification was performed according to European Union (EU) guidelines, using multiple reaction monitoring (MRM) with one precursor and two product ions as identifiers. This method was validated and has been successfully used for routine analysis in the government labs.

Introduction

Nitrofurans antibiotics, furazolidone, nitrofuratoin, furaltadone, and nitrofurazone, have been widely used as food additives for the treatment of gastrointerstinal infections in cattle, pigs, and poultry. Due to mutagenic and genotoxicity, these antibiotics are now banned in food-producing animals. The contamination may not only occur from deliberate and direct misuse of the drugs, but also from contaminated feed and environmental sources. Nitrofuran antibiotics are rapidly metabolized. The metabolites are highly stable with a longer residence time bonded to the meat tissue. Due to the complexity of the sample matrix and no maximum residue limits set, a highly selective and sensitive bioanalytical method is needed by regulatory agencies of many countries.

The LC/MS/MS methods reported in the literature usually have poor sensitivity and reproducibility and time-consuming for sample preparation. We improved the LC/MS/MS method with higher sensitivity and reproducibility and simplified the sample preparation.
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