What question did this study set out to answer?

The central aim is to develop a sustainable analytical method for impurity profiling of nirogacestat using an AQbD approach.

February 13, 2026Open Access

AQbD-driven development of a green RP-HPLC method for nirogacestat with LC–MS structural elucidation

Puntos clave

The central aim is to develop a sustainable analytical method for impurity profiling of nirogacestat using an AQbD approach.
Implemented an AQbD approach to optimize critical method parameters.
Used central composite design to create a robust design space for the HPLC method.
Validated the method according to ICH Q2(R2) guidelines for specificity, linearity, accuracy, precision, and robustness.
Achieved high efficiency in separation with approximately 15,900 plates and excellent symmetry (tailing factor ≤ 1.1).
Demonstrated high specificity and accuracy with recovery rates between 99.4-101.2%.
Showed sensitivity with LOD/LOQ values of 11.58/35.10 ppm for NGT and ≤ 0.52/1.58 ppm for impurities.

Resumen

Nirogacestat (NGT), a recently approved γ-secretase inhibitor for desmoid tumors, requires sensitive and sustainable analytical methods for impurity profiling. This study aimed to develop a green, stability-indicating HPLC method using an Analytical Quality by Design (AQbD) approach. Critical method parameters, including ethanol percentage, ammonium formate buffer concentration, and flow rate, were optimized through risk assessment and a central composite design to establish a robust design space. The final method employed an ethanol-ammonium formate mobile phase (24.14:75.86 v/v, 0.295 mM, pH 3.2) at 1.0 mL/min, achieving baseline separation of NGT, its process impurities, and degradation products with high efficiency (~ 15,900 plates) and symmetry (tailing factor ≤ 1.1). Validation following ICH Q2(R2) confirmed specificity, linearity (R2 > 0.999), accuracy (99.4-101.2% recovery), precision (%RSD ≤ 1.8%), and robustness. Sensitivity was demonstrated with LOD/LOQ values of 11.58/35.10 ppm for NGT and ≤ 0.52/1.58 ppm for impurities. Forced-degradation studies showed 3-14% degradation across stress conditions with mass balance between 95-105%, and peak-purity confirmed the stability-indicating capability. The method's greenness was demonstrated using AGREE, AGSA, CACI, CaFRI, and a multi-assessment tool, with an overall whiteness index of 74%. Overall, this ethanol-based AQbD-optimized method offers a reliable, sustainable, and regulatory-compliant platform for impurity profiling and stability testing of NGT.

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