Simultaneous Determination of Florfenicol and Diclazuril in Compound Powder by RP-HPLC-UV Method

A RP-HPLC-UV method was developed and validated for simultaneous determination of florfenicol and diclazuril in compound powder. The separation involved using a SinoChoom ODS-BP C18(5 μm, 4.6 mm × 250 mm) analytical column. The mobile phase was a mixture of acetonitrile-0.2% phosphoric acid (pH was adjusted to 3.0 with triethylamine). The ratio of acetonitrile and 0.2% phosphoric acid in the mobile phase was 60 : 40 (v/v) from 0 minutes to 6 minutes and 70 : 30 (v/v) from 6.1 minutes to 15 minutes. The flow rate was 1 mL/min. The temperature of the analytical column was maintained at 30°C. The detection was monitored at 225 nm and 277 nm for florfenicol and diclazuril, respectively. The excipients in the compound powder did not interfere with the drug peaks. The calibration curves of florfenicol and diclazuril were fairly linear over the concentration ranges between 50.0–500.0 μg/mL ( r=0.9995 ) and 10.0–100.0 μg/mL ( r=0.9992 ), respectively. The RSD of both the intraday and interday variations was below 2.1% for florfenicol and diclazuril. The method was successfully validated according to International Conference on Harmonisation and proved to be suitable for the simultaneous determination of florfenicol and diclazuril in compound powder.