Development and Validation of Column High-Performance Liquid Chromatographic and Derivative Spectrophotometric Methods for Determination of Levofloxacin and Ornidazole in Combined Dosage Forms

Abstract

Abstract The manuscript describes validated reversed-phase column high-performance liquid chromatographic (RP-HPLC) and first-derivative UV spectrophotometric methods for the estimation of levofloxacin (LFX) and ornidazole (ORNI) in combined dosage forms. The RP-HPLC separation was achieved on a Phenomenex C18 column (250 mm 4.6 mm id, 5 m) using KH2PO4 buffer (pH 6.8)methanolacetonitrile (70 + 15 + 15, v/v/v) mobile phase at a flow rate of 1.5 mL/min and ambient temperature (25 2<sup/>C). Quantification was achieved with photodiode array detection at 295 nm over the concentration range of 110 g/mL for both LFX and ORNI, with mean recovery of 101.7 0.23 and 99.23 1.57, respectively, by the RP-HPLC method. The derivative spectrophotometric method was based on the determination of both the drugs at their respective zero crossing point (ZCP). The first-order derivative spectra were obtained at N =1 (scaling factor), = 2.0 nm (wavelength interval), and the determinations were made at 310 nm (ZCP of ORNI) for LFX and 295 nm (ZCP of LFX) for ORNI over the concentration range of 240 g/mL for both LFX and ORNI. Mean recovery was 99.46 0.96 and 100.9 0.72, respectively, by the first-derivative UV spectrophotometric method. Standard and sample solutions were prepared with methanol as the solvent in both of the methods. These methods were found to be simple, accurate, precise, and sensitive and were applicable for the simultaneous determination of LFX and ORNI in combined dosage forms.