Validation A New RP-HPLC Method For Simultaneous Determination Of Metronidazole And Spiramycin In Bulk Drug And Pharmaceutical Formulations

Abstract

A new RP-HPLC method was developed and validated for Quantitative determination of Spiramycin and Metronidazole in bulk drugs and in pharmaceutical formulations. The separation was done on a CAPCELL PAK C18 column (250 mm x 4.6 mm ID x 5µm), and the detection was performed at 232 nm using UV detector. The mobile phase was a mixture of Methanol–Acetonitrile–Aqueous phosphate buffer (pH 2.64; 0.05 M) (45: 11: 44; v/v) pumped at ambient temperature with a flow rate of  0.9 ml/min, the calibration curve was linear from 5 to 40 µg/ml for both drugs with R² > 0.999. The detection limit (DL) and quantitation limit (QL) were found to be 0.381 and 1.27 µg/ml for Metronidazole and 0.488 and 1.629 µg/ml for Spiramycin, respectively. Accuracy (mean recovery: 100.11, 100.8% for Metronidazole and Spiramycin, respectively) and precision were found to be satisfactory. The optimized method was validated according to The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines [1]. The use of the gradient elution enhanced the separation between the two mentioned drugs in a simple, sensitive, fast and accurate manner within a short analysis time. The proposed method can be used for quality control assay of Spiramycin and Metronidazole in bulk drugs and in pharmaceutical formulations as a result of the ability of the method to separate both drugs from its excipients.