Implementation and Validation of an Analytical Method for Lincomycin Determination in Feathers and Edible Tissues of Broiler Chickens by Liquid Chromatography Tandem Mass Spectrometry

Abstract

Recent studies have detected different antimicrobial residues in broiler chicken feathers, where they persisted for longer periods of time and at greater concentrations than in edible tissues. However, until today, lincomycin behaviour in this nonedible tissue has not been assessed yet. Considering this, an analytical methodology to detect and quantify this antibiotic concentration in feathers, muscle, and liver tissues from broiler chickens was implemented and in-house validated. The methodology will allow the determination of the bioaccumulation of this highly persistent antibiotic in feathers of treated birds. For this purpose, 98% lincomycin and 95% lincomycin D3 standards were used. Methanol was selected as the extraction solvent, and Chromabond® Florisil® cartridges were used for the clean-up stage. The separation of analytes was performed through the analytical column SunFire C18 with a running time of 4 minutes, and the instrumental analysis was performed through an LC-MS/MS, with a liquid chromatograph Agilent® 1290 Infinity, coupled to an AB SCIEX® API 5500 mass spectrometer. An internal protocol for an in-house validation was designed based on recommendations from Commission Decision 2002/657/EC and the Guidance document on the estimation of limit of detection and limit of quantification for measurements in the field of contaminants in feed and food. The average retention time for lincomycin was 2.255 min (for quantifier ion, 126.0). The calibration curves showed a coefficient of determination (r2) greater than 0.99 for all matrices, while recovery levels ranged between 98% and 101%. The limit of detection (LOD) calculated was of 19, 22, and 10 μg·kg−1, and the limit of quantification (LOQ) was of 62, 73, and 34 μg·kg−1 in feathers, muscle, and liver, respectively. This method detects lincomycin in the studied matrices, confidently and accurately, as it is required for designing analytical studies of drug residues in edible and nonedible tissues, such as feathers.

Assessment of various veterinary drug residues in animal originated food products

“…LOD was estimated in range 0.2-0.5 µg/kg and recovery range 82.1-101.4% with relative standard deviation (RSD) <11.1% [3]. Maddaleno et al [4] reported the LOD was 19, 22, and 10 µg/kg and limit of quantification was 62, 73, and 34 µg/kg in feather, muscle, and liver, respectively. Recovery levels reported between 98 and 101% and calibration curve (r²) reported >0.99.…”

“…Maddaleno et al . [ 4 ] reported the LOD was 19, 22, and 10 μg/kg and limit of quantification was 62, 73, and 34 μg/kg in feather, muscle, and liver, respectively. Recovery levels reported between 98 and 101% and calibration curve (r²) reported >0.99.…”

“…Various investigations pointed out the consumption of byproducts from these animals are the major source of human inadvertent antibiotic intake [ 3 ]. Antibacterial drugs have used diverse areas of animal farming as therapeutic agents against different pathogenic microbes [ 4 ]. We all are well known about cow’s milk which contains a good balance of proteins, fats, and carbohydrates.…”

Quantitative Analysis of Spectinomycin and Lincomycin in Poultry Eggs by Accelerated Solvent Extraction Coupled with Gas Chromatography Tandem Mass Spectrometry

“…To date, many methods have been used to measure spectinomycin and lincomycin in animal-derived foods and animal feedstuffs, including fluorescent latex immunoassay (FLI) [11], micellar electrokinetic capillary chromatography combined with ultraviolet detection (MEKC-UVD) [12], enzyme-linked immunosorbent assay (ELISA) [13], high-performance liquid chromatography with electrochemical detection (HPLC-ECD) [14,15], HPLC with fluorescence detection (FLD) [16], HPLC-UVD [17,18], HPLC with evaporative light-scattering detection (ELSD) [19,20], hydrophilic interaction chromatography with mass spectrometry (HILIC-MS) [21], HILIC tandem MS (MS/MS) [22], HPLC-MS [23], HPLC-MS/MS [24][25][26][27][28], gas chromatography-nitrogen phosphorus detection (GC-NPD) [29,30] and GC-MS [30]. The FLI, ELISA, ECD, FLD, UVD and ELSD methods have low sensitivity, specificity, recovery and precision and have many limitations.…”

The vertical transmission of antibiotic residues from parent hens to broilers

“…Consequently, there was a need for matrices in which antibiotics could be detected over a longer time span. For the monitoring of antibiotic use in poultry, the analysis of feathers provided a means to detect antibiotic residues with a long detection window, even after they were excreted fully from muscle and liver (San Martín et al 2007;Cornejo et al 2011b;Berendsen et al 2013;Cornejo et al 2017;Pokrant et al 2018;Církva et al 2019;Maddaleno et al 2019). Using such a noninvasive method, antibiotic use during almost the whole lifespan of a broiler can be monitored, making it possible to better enforce registration of antibiotics use aiming for prudent application (Jansen et al 2016(Jansen et al , 2017.…”