Development and validation of liquid chromatography method for simultaneous determination of multiclass seven antibiotic residues in chicken tissues

Abstract

Background Antibiotics are routinely used on poultry for therapy and prevention of diseases and to enhance animal growth. The objective of this study was to develop and validate a sensitive and reliable liquid chromatography with UV detection (LC-UV) method for the simultaneous determination of seven multiclass antibiotic residues (amoxicillin, ampicillin, penicillin, sulfamethoxazole, gentamicin, ciprofloxacin, and erythromycin) in chicken tissues. Methods The liquid chromatography method with UV detection was optimized for complete separation of the seven selected antibiotic compounds with reversed phase and isocratic elution using Hypersil BDS-C18 (3 µm, 100 mm × 4 mm) column. The mobile phase consisted a ratio of 0.05 M Na2HPO4, acetonitrile and methanol (70:10:20), at UV absorption wavelength of 230 nm. The column thermostat was set at 40 °C, the mobile phase flow rate was 1 mL min−1, and the injection volume was 20 μL. Results All the seven standard compounds were eluted within 14 min. The results for: linearity, precision, sensitivity, accuracy, specificity, decision limit (CCα), detection capability (CCβ), suitability and method robustness were validated according to the criteria of Commission Decision 2002/657/EC guidelines. Calibration plot correlation coefficients ranged from 0.9983 to 0.9998 and the percent relative standard deviations for repeated analysis were below 5% indicating acceptable method precision. The limits of detection (LODs) and quantification (LOQs) ranged from 0.098–0.255 μg kg−1 to 0.297–0.574 μg kg−1, respectively. The accuracy study yielded recoveries in the ranges 98.1–107% for the pure compounds and 94.0–102% for the spiked drug free chicken tissue samples. Conclusions The method was found to be appropriate for simultaneous determination of five different classes of seven antibiotic residues in chicken tissues. Furthermore, this is the first instance for the simultaneous determination of seven multiclass, multi-residues analysis using LC-UV from chicken tissue samples. This is a cost-effective and alternative method with simple instrumentation approach for laboratories that lack highly specialized state-of-the-art instrumentation.

Depletion of Amoxicillin Residue in Edible Tissue of Broiler Chicken by Different Cooking Methods

“…The analytical method used in this study for the determination of Amox in raw, cooked muscles and the surrounding fluids that remained after boiling were prepared, extracted, and analyzed by the previously reported method [ 24 ]. The method was revalidated for this work.…”

Review on Plant-Based Management in Combating Antimicrobial Resistance - Mechanistic Perspective

“…enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay (CLIA), radioimmunoassay (RIA), colloidal gold immunoassay (CGIA) and fluorescence immunoassay (FIA)) ( Ahmad et al, 2017 ; Ahmed et al, 2020b ), as well as biosensor methods have been employed to detect antibiotics residues for food safety and human health. A trend has been increasing for multi-residue detection methods incorporating miniaturization and portability ( Lakew et al, 2022 ).…”

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 Ethiopia, β-lactam antibiotics, i.e., ampicillin, amoxicillin and penicillin are the most widely used class of medicines to treat respiratory tract infections, prostatitis, urinary tract infections, skin, and soft tissue infections of sensitive bacteria in poultry production [11][12][13]. High level of antibiotic residues were detected in meat and milk in studies conducted in Addis Ababa, Bishoftu, and Adama major cities in the country by microbiological or screening methods [14].…”