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Development and Validation of HPTLC Method for Simultaneous Estimation of Gatifloxacin and Ornidazole in Human Plasma

Ambadas R Rote and Prasanna A Kumbhoje

A simple, rapid, sensitive and economic chromatographic method have been developed for simultaneous estimation of gatifloxacin and ornidazole in human plasma by using diclofenac sodium as internal standard The method employes a 20cm x10cm TLC plate precoated with silica gel 60F254 with 250μm thickness on aluminium sheets. The plasma sample was extracted using chloroform:formic acid (4.0:0.4, v/v). The mobile phase used consists of methanol: chloroform: ammonia solution [25%] (8.5: 1: 0.4 v/v/v), having chamber saturation for 20 min at room temperature. The R f values were found to be (R f =0.28±0.00178),(R f =0.77 ±0.00427) and(R f =0.50±0.00337) for GTFX, ORNZ and DICLO respectively. Densitometric analysis was carried out at wavelength 297.86 nm. The linear detector response was observed between 100 ng/band to 600 ng/band for both GTFX and ORNZ. The stability of gatifloxacin and ornidazol in plasma was confirmed during three freeze-thaw cycles (-20 ̊C), on bench during 12 hr and post preparative after 48 hr. The method was validated according to the FDA Bioanalytical guidelines with respect to selectivity, linearity,sensitivity precision and accuracy, recovery and stability.