Abstract
An HPLC method for quantitative estimation of pyrazinamide in rat plasma has been developed. The procedure consists of precipitation of plasma proteins with methanol containing 5-fluorouracil as an internal standard. The mixture was vortexed for 1 min. and centrifuged at 4000 rpm for 10 min. Twenty-five microliters of the supernatant was eluted on μ-Bondapack C
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column at 269 nm. The mobile phase consisted of 2% v/v acetonitrile in 5 mM potassium phosphate buffer at pH 4. The flow-rate was set at 1 ml per min. The retention times for 5-fluorouracil and pyrazinamide were 4 and 9 min. respectively with a detection limit of 0.5 μg/ml for pyrazinamide.