VALIDATION METHODS FOR DETERMINATION OF ENROFLOXACIN LEVEL IN ANIMAL MEDICINE PRODUCTS USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY INSTRUMENT

Abstract

Enrofloxacin is an antibiotic that is often used as a veterinary drug. Analysis of enrofloxacin in veterinary drugs can be carried out using the high-performance liquid chromatography (HPLC) method. The HPLC method was chosen because it is capable of separating the active substance to be measured from other components with high speed and efficiency. This study aims to validate the appropriate analytical method for measuring enrofloxacin levels in veterinary drug products using HPLC. The analytical method was validated using HPLC brand Shimadzu^® 6.1, column C18 150 × 4.6 mm; temperature 30˚C; UV-Vis detector wavelength 278 nm; and a flow rate of 1 mL/min. The mobile phase used was a mixture of 0.05 M NaH₂PO₄ pH 2.5 and acetonitrile with a ratio of 65:35 (volume: volume). The sample was a standard enrofloxacin solution with a concentration of 0.1 μg/mL; 1µg/mL; and 10 μg/mL were analyzed with three replications per sample concentration. The injection volume is 20 µL per injection. The parameters measured were selectivity, linearity, accuracy, precision, limit of detection, and limit of quantification. The results showed good selectivity because the peak area of enrofloxacin was not disturbed by other components. The results of measuring the concentration of standard solutions produce a linear line equation y = 291727x + 37070 with a correlation coefficient value of 0.9999 (r = 0.9999). The accuracy value of each concentration meets the standards ranging from 99.916% to 100.762%. The results of precision calculations are relative standard deviation (RSD) ranging from 5.333% to 10.831%. The detection limit value is 0.003 µg/mL and the quantification limit value is 0.011 µg/mL. The validation test results conclude that the analytical method developed has good validity and can be used to determine enrofloxacin levels in veterinary drug products.