Development and validation of multi-residue and multi-class method for antibacterial substances analysis in non-target feed by liquid chromatography – tandem mass spectrometry

Abstract

A confirmatory HPLC-MS/MS method for the determination of residues of 11 antibacterial substances from different therapeutic class (β-lactams, lincosamides, fluoroquinolones, macrolides, pleuromutilins and sulfonamides) in animal feeds has been developed. The sample preparation is based on extraction with 0.1% formic acid in acetonitrile. Separation of the analytes was performed on biphenyl column with a gradient of 0.1% formic acid in acetonitrile and 0.1% formic acid in Milli-Q water. The developed method was validated following the guidelines included in the European Union Commission Decision 2002/657/EC. Limits of detection ranging from 79.22 to 193.60 µg/kg; instrumental and analytical linearity coefficients were above 0.99 for matrix-match calibration; and relative recoveries ranging from 76.04% to 117.39%. Repeatability of the method was in the range of 2.41-19.76% (CV, %), whereas reproducibility ranged from 6.52 to 28.40% (CV %). The method shown to be efficient and precise for quantification of the 11 antibacterial substances in animal feed. The results demonstrate the feasibility of the method for routine use to monitor these substances in feed. The validated method was successfully applied to eight suspect feed samples collected from the Association of American Feed Control Officials (AFFCO) and feed manufactures from Galicia (Spain) in June and July 2017. Of these 8 non-target feeds, 5 were positive for the presence of tiamulin, tylosin and sulfadiazine.

Determination of Sulfonamides in Feeds by High-Performance Liquid Chromatography after Fluorescamine Precolumn Derivatization

“…These results are in agreement with the data reported by Croubels, who measured sulfadiazine in 27% of feed samples [12]. Patyra et al detected sulfadiazine in three feed samples at concentrations of 250–2960 µg/kg [21]. Kim et al analyzed 156 animal feeds and detected the presence of SAs in feeds that were used on farms, but not the ones that were purchased from markets.…”

“…As described in the literature, good recoveries of SAs from pig, poultry, horse, and cattle feeds have been obtained when extraction was carried out with organic solvents: acetonitrile or methanol with or without water [1,17,19,21,22]. However, the use of methanol or acetonitrile for the extraction of SAs from a feed matrix results in a highly contaminated extract that makes the detection of low levels of SAs impossible.…”

“…The methods of detection and determination of sulfonamides in feed are based on liquid chromatography with UV detector [1], diode array detector [14], and enzyme-linked immunosorbent assay test (ELISA) [10]. Liquid chromatography tandem mass spectrometry (LC-MS/MS or UHPLC-MS/MS) methods have been used for the validation of multi-residue quantification of sulfonamides in feeds [18,19,20,21,22].…”

“…A method for the determination of sulfadiazine and sulfamethazine in feed was developed by Patyra et al, who used an LC-MS/MS instrument. Average recovery was in the range of 76.0–77.4%, and LOQ values ranged from 106.2–174.6 µg/kg [21].…”

Innovative process methodologies for the agro-food sector: reorganization of technical processes to acquire new skills

“…The reasons that have led the research team to consider the above critical issues arise from the observation, in recent years, of organizational difficulties, problems in quality control, possible delays in response times to customers and, as a result, in the potential loss of customers as well as the lack of opportunity to provide innovative services to each type of customer. Multi-residual analysis is aimed at identifying residual substances on products intended for food in the form of any matrix: solid, liquid or gaseous [6][7][8]. They are particularly requested and widespread on products of vegetable, animal and water origin.…”