Development and Validation of HPTLC Method for Determination of Ofloxacin in Human Plasma

To develop a simple, selective and sensitive high performance thin layer chromatographic method for the determination of Ofloxacin in human plasma. The method had been validated for linearity, precision, accuracy and stability following EMEA and US CDER guidelines for bioanalytical method validation. Sample was prepared by liquid – liquid extraction using chloroform. Chloroform layer seperated was evaporated and reconstituted in methanol and spotted on TLC plates precoated with silica gel 60 F254. Mirtazepine was used as an internal standard. The mobile phase consisted of a mixture of n-butanol: ethanol: ammonia in tha ratio of 5:5:4 v/v/v. The drug showed considerable absorbance at 294 nm. The method was found to be linear over the concentration range of 50–600 ng/band. Mean drug recovery was found to be 98.76%. Ofloxacin in plasma samples was stable parameters as per EMEA and US CDER guidelines. The method was found to be precise, accurate and can further be extented to pharmacokinetic studies for therapeutic drug monitoring of Ofloxacin in routine clinical practices.