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Bioanalytical method development and validation of bupropion hydrochloride in rat plasma by RP-HPLCKeywords: Bupropion HCl , HPLC , Bioanalysis , Rat plasma , UV detection. Abstract: A novel, rapid, sensitive, accurate and specific HPLC assay with UV-Visible detection (250 nm) was developed and validated for the determination of bupropion hydrochloride in rat plasma. Phenacetin was used as internal standard (IS). The plasma proteins were precipitated by a single step protein precipitation extraction procedure using methanol (acidic pH). Chromatographic separation was achieved with a combination of acetonitrile and 0.01 M potassium dihydrogen phosphate (pH 3.0 adjusted with orthophosphoric acid) in the gradient mode on a C18 (250 mm × 4.6 mm, 5 μm) analytical column. Mean recovery of bupropion hydrochloride from rat plasma was around 55 % for 2.5-50 μg/ml concentrations. The assay exhibited good linear relationship with an r2 of 0.9999. Lower limit of Quantification limit (LLOQ) was 1.84 μg/ml of bupropion hydrochloride and accuracy and precision were over the concentration range 2.5-50 μg/ml. The method was validated with good sensitivity, accuracy, precision and recovery. The assay can be applied successfully to pharmacokinetic studies.
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