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

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.

Determination of Colistin B in Chicken Muscle and Egg Using Ultra-High-Performance Liquid Chromatography–Tandem Mass Spectrometry

“…It was further modified after studying the core literature using LC–MS/MS for polypeptide antibiotics determination in animal origin food. The published literature for sample preparation primarily discusses the deproteinization step (referred to as acidic pretreatment), performed with the help of organic solvents (acetonitrile and methanol) and acids (trichloroacetic acid and hydrochloric acid) in varied combinations, as shown in Table 3 [ 2 , 9 , 10 , 11 , 12 , 16 , 17 ].…”

“…Colistin is a cationic peptide antibiotic with weak acid (carboxylic acid) and base (amino) functional groups. Bladek et al [ 2 ] have reported that colistin could be found in three forms—deprotonated base form, neutral form or protonated acid form. Colistin is highly polar in nature, hence why in the present study polar extraction solvents i.e., methanol and acidified water, were used [ 18 ].…”

“…Colistin (polymyxin E) is a cationic peptide non-ribosomal antibiotic synthesized by Bacillus polymyxa subspecies colistinus, and is an active member of the polymyxin group [ 1 , 2 ]. Colistin is stated to be composed of approximately thirty different constituents.…”

“…Colistin is stated to be composed of approximately thirty different constituents. Out of all the components, colistin A (polymyxin E1) and colistin B (polymyxin E2) are the most predominant, accounting for ≥85% of colistin utilized in pharmaceutical products, which only differ because of the single carbon in the fatty acyl moiety [ 2 ]. Owing to high toxicity, this peptide antibiotic has been banned for human consumption, but still it is extensively used as a feed additive and veterinary drug in animal husbandry [ 3 ].…”

“…[2,[9][10][11][12]16,17].arison of earlier reported sample preparation protocols used for colistin B detection with newly proposed…”