Liquid Chromatography-Tandem Mass Spectrometry Determination and Depletion Profile of Chlortetracycline, Doxycycline, and Oxytetracycline in Broiler Chicken Muscle After Oral Administration

Abstract

A simple and fast method based on liquid chromatography–electrospray ionization-tandem mass spectrometry (LC–ESI-MS/MS) was developed and validated for determination of tetracyclines in broiler chicken muscle. Sample preparation was performed using extraction with acetonitrile, followed by low-temperature purification (at − 20 °C) and further concentration. The chromatographic analysis was carried out using a Zorbax SB-C18 column with gradient elution using water and methanol both acidified with 0.025 M of formic acid. Mass spectrometry with electrospray ionization was operated in positive polarity using selected reaction monitoring (SRM) analysis mode, achieving the requirements of four identification points for each compound. Demeclocycline was used as internal standard. The method validation was done according to the criteria of Commission Decision 2002/657/EC. Parameters such as recovery, matrix effects, selectivity/specificity, linearity, precision (intra- and inter-day precision), accuracy, limits of detection (LOD) and quantification (LOQ), decision limit (CCα), detection capability (CCβ), and robustness were determined. Intra-day precision values were within the range 2.2–5.8% and inter-day precision was less than 10% for all analytes. Accuracy ranged from 98.2 to 103.2%. The method was successfully applied for depletion studies of chlortetracycline, doxycycline, and oxytetracycline in broiler chicken tissues after multiple oral administrations. After the depletion studies, the present study support more prudent use of CTC, DOX, and OTC for treatment of chickens and suggest a dose of 60 mg kg−1 body weight for CTC and OTC and 20 mg kg−1 body weight for DOX, orally administrated for five consecutive days.