A Quality by Design-Driven
Stability indicating UPLC Method for the Development and Validation of Simultaneous
Nirogacestat and Impurity Quantification
Umamaheswara Rao K.V. and Shorgar Neetu
Res. J. Chem. Environ.; Vol. 29(8); 54-65;
doi: https://doi.org/10.25303/298rjce054065; (2025)
Abstract
The present study intended to propose a robust and reliable UPLC method for the
simultaneous quantification of Nirogacestat and its impurities 1 to 5. A systematic
quality by design (QbD) approach was employed to optimize critical method parameters
that ensure enhanced resolution, peak symmetry and sensitivity. The chromatographic
conditions were carefully selected based on an extensive literature review and theoretical
physicochemical assessments. Various column chemistries and mobile phase compositions
were evaluated to achieve optimal separation of Nirogacestat and its impurities.
A total of 13 experimental trials were conducted and the results were analyzed through
perturbation charts, contour plots and 3D surface plots to determine optimal chromatographic
conditions. Finally, the optimized conditions comprise of an X-Bridge C18 (50 mm
× 4.6 mm, 2.1 μm) column, acetonitrile and 0.1% trifluoroacetic acid in a 40:60
(v/v) ratio at 0.3 mL/min flow as mobile phase and 299 nm wavelength. This method
exhibits excellent linearity over the studied concentration ranges, with correlation
coefficients (R²) greater than 0.999 for Nirogacestat (50–300 μg/mL) and its impurities
(2.5–15.0 μg/mL).
The method demonstrated high accuracy with mean recovery rates of 98-102 % for Nirogacestat
and its impurities across three concentration levels. Sensitivity assessment reveals
sensitive detection and quantification limits for Nirogacestat and its impurities
respectively ensures reliable detection of trace impurities. The robustness study
indicates that minor variations in flow rate and organic modifier composition had
minimal impact on chromatographic performance. Forced degradation study confirms
the method's stability-indicating nature. The purity angle and purity threshold
assessment in stress study confirm the absence of co-elution proving the method’s
capability for impurity profiling. In conclusion, this study establishes a novel,
precise and reliable chromatographic method for the simultaneous determination of
Nirogacestat and its impurities for routine pharmaceutical analysis and quality
control applications.
