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

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 NaHPO, 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, 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 to 0.297-0.574 μg kg, 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.