Stability Indicating HPLC Method for Quantitative Estimation of Pralsetinib in Pralsetinib Solid Dosage Form

Introduction: Pralsetinib is an oral receptor tyrosine kinase inhibitor. Pralsetinib, sold under the brand name Gavreto, is a medication approved for RET mutation-positive medullary thyroid cancer (MTC) and RET fusion-positive differentiated thyroid cancer (DTC) refractory to radioactive iodine (RAI) therapy. Objectives: An analytical method has been developed for estimation of Pralsetinib in solid dosage form by using isocratic reverse phase high performance liquid chromatography assay method. The present work describes a simple, stability indicating HPLC method for the determination of Pralsetinib in Pralsetinib in solid dosage form according to ICH guidelines Methods: Samples are analysed by using stationary phase Agilent Zorbax SB-C18 (150 x 4.6 mm, 5µ) and the mobile phase consists of pH 4.5 ammonium acetate buffer and methanol in the ratio of (65:35 volume/volume). The column temperature was maintained at 40°C and sample cooler temperature was maintained at 5°C, injection volume 10 µL, flow rate 1.0 mL/min and wavelength 215 nm, run time 25 minutes. The retention time of Pralsetinib was noted to be 6.45 min respectively. Results:A precise, accurate and selective stability-indicating reverse phase high performance liquid chromatographic assay method has been developed for the quantitative estimation of Pralsetinib in Pralsetinib in solid dosage form. As there is no interference of blank and placebo at the retention time of Pralsetinib analyte peak. Degradation study results were shown No degradation was observed for the solid state and liquid state under the investigated stress conditions. Hence it can be concluded that Pralsetinib is stable product. In order to obtain system precision, a study was conducted with six replicate injections. %RSD was estimated from the peak areas of Pralsetinib found to be 0.42% respectively. The acceptance limit would be not greater than 2.0, and the results were found to be within the acceptance limits. The method precision of test method was evaluated by preparing six samples of Pralsetinib samples as per test method. The average content of the six preparations and % RSD for the six observations were calculated. Relative standard deviation for method precision was found to be 0.26%. The acceptance limit should be not more than 2.0 and the results were found to be within the acceptance limits. The proposed HPLC method was linear over the range of 101.44-304.32 µg/mL for Pralsetinib, the correlation coefficient was found to be 0.9999. The accuracy studies were shown as % recovery for Pralsetinib 50% to150% level. The limit of % recovered shown is in the range of 98 and 102% and the results obtained were found to be within the limits. Hence the method was found to be accurate. Solution stability of standard and sample solutions at different time intervals studied, from the above results, it is concluded that standard solution is stable upto 48 hours at bench top and in refrigerator condition (2-8°C). Sample solutions are stable up to 48 hours in both the conditions bench top (RT) and refrigerator (2-8°C). The method is robust for changes like flow rate, column oven temperature, pH variation and organic phase of mobile phase. Conclusions: The developed method was validated for various parameters as per ICH guidelines like specificity, system suitability, precision, linearity, accuracy, solution stability and robustness. The results obtained were within the acceptance criteria. So, it can be concluded that the developed method is simple, precise, cost-effective, eco-friendly, safe and can be successfully employed for the routine analysis of Pralsetinib in bulk and pharmaceutical dosage forms. The method was validated as per ICH guidelines. The proposed method was found to be accurate, reproducible and consistent.