The present work highlights the spectrophotometric method development and validation for the assay of ampicillin, a broad-spectrum β-lactam antibiotic. Two methods have been proposed by employing popular chromogenic reagents viz 3-methyl 2-benzothiazolinone hydrazone (MBTH) and para dimethyl amino cinnamaldehyde (PDAC). Method A involves the coupling of ampicillin with MBTH in the presence of ferric chloride through the oxidative mechanism, resulting in the formation of green coloured solution with a maximum absorption peak at 620 nm. In method B, the condensation reaction of ampicillin with an acidic solution of PDAC gave a coloured reddish-brown complex, showing a maximum absorption peak at 514nm. Linear calibration curve for MBTH and PDAC has been obtained while obeying Beer’s law in the concentration range of 0.5 -1 and 5-15 μg/ml having a good correlation coefficient of 0.9985 and 0.9974 respectively. The sensitivity of the submitted methods was found to be more than the reported methods. Both methods were successfully applied to a pharmaceutical formulation for quantifying the ampicillin effectively. Therefore, the developed spectrophotometric methods can serve as a good analytical assay procedure for ampicillin to undergo quality control in pharmaceutical setups.
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