Design of experiment based validated stability indicating RP-HPLC method of temozolomide in bulk and pharmaceutical dosage forms

A simple, precise, stability indicating RP-HPLC method was developed and validated for determination of Temozolomide (TMZ) in bulk sample and nanostructured lipid carriers. A Box-Behnken statistical design with three factors and three levels was employed to optimize the chromatographic conditions. The separation was achieved using solvent system combination using methanol: water (30:70) with acetic acid (0.25%). The developed method was validated for linearity, precision, accuracy, limit of detection and limit of quantitation, and robustness in accordance with the ICH guidelines. Further degradation study of TMZ in different stress conditions was studied. The method was found linear with regression co-efficient of 0.999 over a wide range of 10-100 µg/mL. All the validation parameters were found to be within the acceptance range. The developed method was successfully applied to estimate the amount of TMZ in pharmaceutical formulations.