Determination of Polypeptide Antibiotic Residues in Food of Animal Origin by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry.

Tomasz Bladek,Andrzej Posyniak,Iwona Szymanek-Bany

doi:10.3390/molecules25143261

Tomasz Bladek, Andrzej Posyniak + Show 1 more

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https://doi.org/10.3390/molecules25143261

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Abstract

A novel UHPLC-MS/MS method for the determination of polypeptide antibiotic residues in animal muscle, milk, and eggs was developed and validated. Bacitracin A, colistin A, colistin B, polymyxin B1, and polymyxin B2 were extracted from the samples with a mixture of acetonitrile/water/ammonia solution 25%, 80/10/10 (v/v/v), and put through further evaporation, reconstitution, and filtration steps. The chromatographic separation was performed on a C18 column in gradient elution mode. Mass spectral acquisitions were performed in selective multiple reaction monitoring mode by a triple quadrupole mass spectrometer. The method was validated according to the criteria of Commission Decision 2002/657/EC. The method quantifies polypeptides in a linear range from 10 to 1000 μg kg−1, where the lowest concentration on the calibration curve refers to the limit of quantification (LOQ). The recoveries ranged from 70 to 99%, the repeatability was below 13%, and within-laboratory reproducibility was lower than 15%. The decision limit (CCα) and detection capability (CCβ) values were calculated, and ruggedness and stability studies were performed, to fulfill the criteria for confirmatory methods. Moreover, the developed method may also be used for screening purposes by its labor efficiency.

Highlights

Polypeptide antibiotics are a group of antimicrobials with a variety of actions against manyGram-negative and Gram-positive bacteria
LC-MS/MS assays tend to have a lower sensitivity for peptides than for small of fragmentation
The mass parameters were optimized by infusing standard solutions of polypeptide fragmentation

Summary

Introduction

Polypeptide antibiotics are a group of antimicrobials with a variety of actions against manyGram-negative and Gram-positive bacteria. Polypeptide antibiotics are a group of antimicrobials with a variety of actions against many. Members of the polypeptide family are bacitracin, colistin. A, colistin B, polymyxin B1, and polymyxin B2. These large-molecular-mass compounds have a common structure of a heptapeptide ring with a polypeptide side chain (Figure 1). Bacitracin is produced by Bacillus licheniformis and Bacillus subtilis and is a mixture of several closely related polypeptides, mainly consisting of bacitracin A (above 50%), and of bacitracin B1, B2, C, and F to a lower extent [1]. Colistin ( known as polymyxin E) is an important member of the polymyxin group of cationic peptide antibiotics and is produced by cultures of Bacillus polymyxa var. At least thirty different components have been found in commercially available colistin

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