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

Abstract

A simple, rapid, sensitive, and economic chromatog. method were developed for simultaneous detn. of gatifloxacin and ornidazole in human plasma by diclofenac sodium as internal std. The method employes a 20cm × 10cm TLC plate precoated with silica gel 60F254 with 250 μm thickness on aluminum sheets. The plasma sample was extd. using chloroform:formic acid (4.0:0.4, vol./vol.). The mobile phase used consists of methanol:chloroform:ammonia soln. [25%] (8.5:1:0.4 vol./vol./v), having chamber satn. for 20 min at room temp. The Rf values were found to be (Rf = 0.28 ± 0.00178), (Rf = 0.77 ± 0.00427) and (Rf = 0.50 ± 0.00337) for GTFX, ORNZ and DICLO resp. Densitometric anal. was carried out at wavelength 297.86 nm. The linear detector response was obsd. between 100 ng/band to 600 ng/band for both GTFX and ORNZ. The stability of gatifloxacin and ornidazole in plasma was confirmed during 3 freeze-thaw cycles (-20°C), on bench during 12 h and post preparative after 48 h. The method was validated according to the FDA bioanal. guidelines with respect to selectivity, linearity, sensitivity precision and accuracy, recovery, and stability. [on SciFinder(R)]