Green metrics and quality by design tool evaluated UPLC content determination method for Sparsentan in formulation sample

• UPLC method developed with economical run time of 3 mins. • The greenness of the method underwent thorough investigation and validation through the utilization of four distinct metrics: analytical eco-scales, NEMI, GAPI, and AGREE. • The strength of the approach was evaluated by utilizing the QbD statistical tool alongside DoE, revealing that the technique showed durability and adaptability. Sparsentan (SRN) is an endothelin and angiotensin II receptor antagonist indicated to reduce proteinuria in adults with primary immunoglobulin a nephropathy at task of rapid disease progression. In this study, we developed and validated a method to determine SRN in pharmaceutical dosage form using a reverse phase-UPLC technique. The specificity of the method was achieved in analytical column ACQUITY UPLC BEH C18 analytical column [1.7 µm, 2.1 mm X 150 mm] using 0.1% Trifluoroacetic acid (pH 4.0) and Methanol in the ratio of 65:35 v/v as a mobile phase. The flow rate was 0.3 mL/min. The column temperature was 42°C and the sample compartment temperature was 5°C. The injection volume was 1 µL, detection was carried out at 263 nm in UV, and the total run time was 3.0 minutes. The method was found to be linear from LOQ to 150 % concentration level, the correlation coefficient value found 0.9995. The recovery was performed from 50% to 150% level concentration and mean recovery was found 99.19 % to 100.54%. The method precision data resulted 0.3%. The samples were made for forced degradation under hydrolysis, oxidation, thermal and photolytic conditions. Method robustness studied with QbD tool. The degradation and validated study results indicate its stable nature. The method was validated according to the ICH guidelines and found to be specific, linear, rugged, robust and accurate. Method greenness studied with advanced metrics GAPI, NEMI, Eco-scale and AGREE.