Development and Validation of RP - HPLC Method for the Determination of Azathioprine in Bulk and Pharmaceutical Dosage form

Existing literature extensively documents numerous scholarly studies on azathioprine. The primary aim of this investigation was to come up with a straightforward, quick, and precise RP-HPLC methodology capable of measuring azathioprine accurately in pharmaceutical dosage forms. Column Agilent Zorbax Bonus-RP-C18 (250 x 4.6mm, 5µm) served as the stationary phase for the chromatographic separation, while a mobile phase consisting of ACN and 0.1% TFA in water (45:55) was administered isocratically at an elution rate of 1ml/min. The detection was executed at 278nm employing a UV detector. Furthermore, we conducted statistical validation for the linearity, exactness, precision, system suitability, and specificity of the approach in both bulk drug and pharmaceutical formulations for the quantitative analysis of azathioprine. The developed methodology exhibited remarkable attributes, demonstrating rapidity with a retention time of 2.67 minutes. It has revealed exceptional accuracy, achieving a mean recovery rate ranging from 99.92% to 100.06%, and displayed remarkable precision, with a relative standard deviation for the system of less than 2%. Moreover, the approach demonstrated a linear relationship between the concentrations of 40 to 60µg/ml, with a significant coefficient of correlation of 0.995. Furthermore, we observed detection and quantification limits of 6.17µg/ml and 18.69µg/ml, correspondingly. Notably, the anticipated approach proved to be straightforward, rapid, exact, accurate, and reproducible, making it suitable for the accurate quantification of azathioprine in both pure drug samples and tablet formulations.