In-House Validation of HPLC-MS/MS Methods for Detection and Quantification of Tetracyclines in Edible Tissues and Feathers of Broiler Chickens

Abstract

For the detection of tetraciclines in feathers, muscle and liver, an internal protocol was designed for in-house validation of two methods, through high performance liquid chromatography with tandem mass spectrometric (HPLC-MS/MS) techniques. This protocol was based on the recommendation of the decision 2002/657/EC from the European Community as well as the food and drug administration (FDA) VICHGL49. Limit of detection (LOD) was set at 20 μg kg -1 and the limit of quantification (LOQ) ranged from 21.5 to 24.2, 21.2 to 21.6, and 25.0 to 27.7 μg kg -1 in feahers, muscle and liver samples, respectively, for all analytes. The calibrations curves show a coefficient of determination (R 2 ) above 0.98, 0.99 and 0.96 for feathers, muscle and liver samples, respectively. Analyte recovery ranged from 92 to 108%. In conclusion, these methods can be deemed accurate and reliable, and their validation is a fundamental step to be performed in depletion studies on these matrices.

Concurrent Determination of Tigecycline, Tetracyclines and Their 4-Epimer Derivatives in Chicken Muscle Isolated from a Reversed-Phase Chromatography System Using Tandem Mass Spectrometry

“…Therefore, methanol was used to dissolve the standard in the preparation of the stock solution, a slightly acidic solvent was used as the diluent in the preparation of the working solution, and trichloroacetic acid was used in both diluent and mobile phase B to reduce possible solvent interference. Acidified protein precipitants (methanol and acetonitrile) [ 41 , 42 , 47 , 49 ] and buffer solution [ 43 , 44 , 45 , 46 , 48 , 50 ] were used as extractants, and acidifiers included formic acid, acetic acid, trifluoroacetic acid and EDTA. EDTA was used as the extractant in this study to minimize the chelation of tetracyclines with metal cations in biological samples while maintaining the freeness of the targets [ 41 ].…”

“…The analytes in this study included four tetracyclines (TC, CTC, OTC and DOXY), as specified in China’s current national food safety standards, coupled with their three 4-epimer derivatives (4-epi-TC, 4-epi-CTC and 4-epi-OTC) and the novel glycylcycline (TGC). Of the studies where HPLC–MS/MS was used as the detection method, some studies [ 41 , 42 , 43 , 44 , 45 , 46 ] investigated a range of analytes that was not as comprehensive as that used in the present study. The analytes involved in the study by Nakazawa et al [ 48 ] included demeclocycline but did not contain 4-epimer derivatives or TGC.…”

Trace level enrichment and clean-up of β-lactam antibiotic residues in edible chicken tissues and feathers by solid phase extraction for quantitative determination utilizing liquid chromatography

“…In order to provide safe food and secure consumer health, antibiotic usage within the developed countries officially monitored through the materials collected on post mortem, such as muscles, liver, skin, fat in the food of animal origin using the established maximum residue limits (MRLs). While these limits have been set for most antibiotics for food producing animals, there are no regulatory limits specified MRLs for poultry feathers, as these structures are not used for direct consumption [7]. However, recent studies have detected different antimicrobial residues in animal by-products such as broiler chicken feathers.…”

“…However, recent studies have detected different antimicrobial residues in animal by-products such as broiler chicken feathers. The researchers have found higher concentrations of antimicrobial residues in feathers than in edible tissue samples and they were persisted for longer periods [7][8][9][10].…”

Development and application of a novel electrochemical immunosensor for tetracycline screening in honey using a fully integrated electrochemical Bio-MEMS

“…Several analytical methods currently exist for TC detection and quantification. High performance liquid chromatography coupled to mass spectrometry (Pokrant et al, 2018), capillary electrophoresis coupled with electrochemiluminescence (Deng et al, 2012), enzyme immunoassay (Gaurav et al, 2014), liquid chromatography-mass spectroscopy (LC-MS) (Desmarchelier et al, 2018) have been described for confirmatory analysis. Other methodologies based on spectroscopy analysis are described (Qin et al, 2016); unfortunately, they suffer from a lack of sensitivity compared to chromatographic techniques (Mitra, 2004).…”