Stability indicating methods for the determination of norfloxacin in mixture with tinidazole

Three stability indicating assay methods are developed for the determination of norfloxacin (Nor) in the presence of its decarboxylated degradation product and in mixture with tinidazole (Tnd). The proposed methods are reversed phase ion pair liquid chromatography (LC), thin layer densitometry (TLC) and second derivative ratio spectra zero crossing spectrophotometry ((2)DD). Chromatographic separation was achieved on mu-Bondapack C18 column 5 microm (300 mm x 3.9 mm, I.D.) and precoated silica gel TLC stationary phases for LC and TLC methods, respectively. Mobile phases consisting of phosphate buffer pH 3.2 : methanol (3 : 1, v/v) containing 0.005 M pentane sulfonic acid sodium salt and isopropanol : butanol : concentrated ammonia : water (25 : 50 : 5 : 25, v/v/v/v) were used for resolution of Nor and Tnd by both techniques, respectively. Detection was carried at 280 nm. In the ratio spectra method, detection of Nor was carried at 282 nm. Linearity, accuracy and precision were found to be acceptable over concentration ranges of 20-225 microg/ml, 0.8-4 microg/spot and 1-7 microg/ml for Nor by LC, TLC and (2)DD methods and over concentration ranges of 37.5-375 microg/ml and 4.8-20 microg/spot for Tnd by LC and TLC methods respectively. The suggested methods were successfully applied for the determination of both drugs in bulk powder, laboratory prepared mixtures and in commercial samples. Statistical comparison between the results obtained by the proposed and the reference methods was carried out using Student t-test, F ratio and one way ANOVA.