Original Articles
Bhavin K. Patel and Rajesh H.
Abstract
A simple, sensitive and specific reverse phase - high performance liquid chromatography (RP-HPLC) method have been developed and validated for the estimation of Temozolomide (TMZ) in its plasma and brain homogenate of Rats. TMZ concentration in plasma and brain homogenate was determined on a C18 column after liquid-liquid extraction. Elution was applied with the mixture of 0.1% aq. Acetic Acid- Acetonitrile (90:10 V/V). To prevent chemical degradation of TMZ, plasma samples were acidified to pH < 3. The flow rate was 1 ml/min and separation was monitored by UV detection at 316 nm. Chromatogram showed peak at a retention time of 5.643± 0.0811 and 5.781±0.0545 for the plasma and brain respectively. All validation parameters met the acceptance criteria. Calibration curve, prepared using freshly spiked plasma and brain homogenate samples,was linear within the range of 1μg/ml to 100 μg/ml. Validation of the method for linearity and range, intra-day precision, selectivity, specificity and limits of detection and quantification were obtained. The method was found to be sufficiently accurate and precise over the studied range of concentrations. The developed method is suitable for pharmacokinetic studies in rats.