New Analytical Method Development and Validation of Ciprofloxacin and Ornidazole in Human Plasma by High Performance Thin Layer Chromatography

Abstract

Objective: The objective of the method was to develop a simple, rapid, sensitive, selective and economic high performance thin layer chromatographic method for simultaneous determination of ciprofloxacin and ornidazole in human plasma by using tinidazole as an internal standard. Method: The plasma sample was extracted using methanol: formic acid (5.5:0.5 v/v) and known amount of extract was spotted on precoated silica gel 60 F 254 plates using Camag Linomat V auto sampler. A concentration range from 100–700 ng/spot for both drugs was used for calibration curve. The percent recoveries of Ciprofloxacin and ornidazole were found to be 81.02 to 86.26 and 79.73 to 82.16 respectively. The mobile phase used consists of chloroform: methanol: triethylamine (9.0: 0.8: 0.4 v/v/v). Densitometric analysis was carried at wavelength 291 nm. Result: The R f values for ciprofloxacin, ornidazole and tinidazole were found to be 0.18 ± 0.057, 0.49 ± 0.0.0057 and 0.75 ± 0.0054 respectively. The stability of ciprofloxacin and ornidazole in plasma were confirmed during three freeze-thaw cycles (-20oC), on bench during 12 h and post preparative stability study. The proposed method was validated statistically by performing recovery study for determination of ciprofloxacin and ornidazole in human plasma. Conclusion: The proposed method was found to be a simple, rapid, sensitive, selective and economic high performance thin layer chromatographic method for simultaneous determination of ciprofloxacin and ornidazole in human plasma In future this method can be used for clinical and pharmacokinetic studies. Key words: Ciprofloxacin, Ornidazole, HPTLC, Human plasma, Liquidliquid extraction.