ABSTRACT:
The present work was aimed at establishing a rapid, specific and simple high performance liquid chromatography (HPLC) method for quantitative estimation of allopurinol (AL) in mice serum and kidney homogenate. Serum and kidney homogenate samples were prepared using protein precipitation method. The prepared samples were then analyzed using Agilent’s ZORBAX, SB C18 column, photodiode array detector and retention time observed was 1.6 min for allopurinol. Further, the HPLC method was validated in order to demonstrate its linearity, accuracy, robustness, precision, specificity, limit of detection (LOD) and limit of quantification (LOQ). The linearity of calibration curve was observed in the range of 0.5-12.5 µg/ml and 0.5-5 µg/ml for kidney homogenate and serum, respectively. The LOD for serum and kidney samples was obtained as 63 ng/ml and 241 ng/ml, respectively and LOQ for serum and kidney samples was obtained as 192 ng/ml and 730 ng/ml, respectively. The accuracy, precision and robustness were found to be in compliance with ICH guidelines.