Development and validation of Apalutamide related substances method in solid dosage forms using HPLC.

Anti-cancer drug formulation Apalutamide 60mg tablets related substances method was developed and validated by using liquid chromatography gradient elution method. All the impurities and degradants were separated by using the Luna Omega 5μm Polar C18, (250 x 4.6) mm HPLC column with a 1.0 mLmin -1 flow rate. The detection was done at 225 nm by injecting the 10 μL of injection volume by controlling the sample temperature at 10 °C and maintaining the column compartment temperature at 30 °C. The total run time was 85 mins. 0.01M Di sodium Phosphate dihydrate pH 4.20 ± 0.05 buffer mixed with acetonitrile in the ratio of 73:27 (v/v) used as mobile phase A. Mobile phase B consists of water and acetonitrile in the ratio of 30:70 (v/v). The proposed method was validated as per the current regulatory guidelines. The method precision at 100 % specification level % RSD results were found 1.41, 1.74, 1.84, and 1.66 for all four impurities. The accuracy results were obtained between 96.0 % to 106.3 % for LOQ to 150 % level. The standard and sample solutions stability were established for 44 hrs at 10 °C. The correlation coefficient (r) value is > 0.999 found for all the four impurities, indicating good linearity between concentration and peak response. 0.9999, 0.9999, 0.9999, and 1.0000 these results express the method linearity. Available three filter compatibility was proven and concluded that 0.45μm Nylon, PTFE, and PVDF filters are suitable. The robustness of the method was established by varying all the conditions. The method specificity was proven and the forced degradation data reveals the method's stability-indicating nature.