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 (R2) 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.
Highlights
Antimicrobials are the main therapeutic tool currently being used to control infectious pathologies from bacterial origin, they are used to improve the efficiency of farming systems of animals destined for human consumption
Several institutions and organisms have established directives for the validation of analytical methods such as the Codex Alimentarius by FAO, VICH GL 49 by the Food and Drug Administration (FDA)[22] and the 2002/657/CE decision by the European Commission.[23]. Based on these international guidelines, this study developed an internal protocol to validate analytical methodologies of HPLC coupled to mass spectroscopy detection (HPLC-MS/MS)
The analytical methods developed in this work for feathers, muscle and liver samples were validated against an internal protocol
Summary
Antimicrobials are the main therapeutic tool currently being used to control infectious pathologies from bacterial origin, they are used to improve the efficiency of farming systems of animals destined for human consumption. For example, tetracyclines are widely used due to its broad spectrum activity These antimicrobials are naturally synthesized by Streptomyces spp. and were discovered in 1948 by Benjamin Duggar. They are active against Gram positive and Gram negative bacteria, either aerobic or anaerobic, as well as Spirochaetes, Actinomyces, Rickettsia, Chlamydia, Mycoplasma and even some protozoa. Such a broad spectrum makes them especially valuable for the treatment of respiratory and digestive diseases.[1,2]. They are best absorbed during fasting and worst when ingested concurrently with calcium salts or aluminium hydroxide
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