Novel Separation and Quantitative Determination of Levofloxacin, Prulifloxacin, Gatifloxacin, Sparfloxacin, Moxifloxacin and Balofloxacin Fluoroquinolone Antibacterials in Pharmaceutical Dosage Forms by RP-HPLC Method

For the first time, Simple, selective, sensitive RP-HPLC method developed for the separation and
quantitative estimation of Levofloxacin (LEVO), Prulifloxacin (PRFX), Gatifloxacin (GATI),
Sparfloxacin (SPAR), Moxifloxacin (MOXI) and Balofloxacin (BALO) relating to fluoroquinolone
antibacterials in pharmaceutical dosage forms. The important advantage of the developed method
was that the six separate drugs could be estimated on a single chromatographic system without
modifications in detection wavelength and mobile phase by RP-HPLC. The chromatographic
separation of the selected drugs carried out on Welchrom C18 column consisting of 250 mm X4.6 mm,
5 μm particle size utilizing mixture of 10 mM phosphate buffer (pH 3.1): Acetonitrile in the ratio of
70:30,v/v as mobile phase at the flow rate of 1 mL/min with detection wavelength at 293 nm by using
UV spectrophotometric detector with total run time of 10 minutes and 3.613, 4.230, 4.707, 5.497,
5.880 and 6.253 minutes of retention time, 12,261, 12,554, 13,157, 14,761, 14,912 and 15,916 of
plate number, 1.106, 1.067, 1.040 and 1.073, 1.030 and 1.086 tailing factors were obtained for LEVO,
PRFX, GATI, SPAR, MOXI and BALO respectively. All calibration curves for six drugs showed
indicated linearity over a concentration range of 2-10 μg/mL. The results regarding to limit of detection
(LOD) and limit of quantitation (LOQ) for LEVO, PRFX, GATI, SPAR, MOXI and BALO were found to
be 0.116 μg/mL and 0.348 μg/mL; 0.152 μg/mL and 0.460 μg/mL; 0.084 μg/mL and 0.255 μg/mL;
0.186 μg/mL and 0.558 μg/mL, 0.162 and 0.493, 0.112 and 0.390 respectively. These results
clearly show the low values of LOD and LOQ. The said proposed method utilized for quantification
of the marketed formulation. The mean assay values for LEVO, PRFX, GATI, SPAR, MOXI and BALO
arrived at 99.317±0.990%, 99.9±0.04%, 99.9±0.02%, 99.45±0.01%, 99.945±0.056% and
99.68±0.09% respectively.