Analytical Quality by Design for Chiral Pharmaceuticals: A Robust HPLC Method for Upadacitinib Enantiomeric Quantification

Abstract

Ensuring the enantiomeric purity of chiral pharmaceuticals is paramount for patient safety and therapeutic efficacy. Upadacitinib (UPA), a vital Janus kinase 1 (JAK-1) inhibitor for rheumatoid arthritis treatment, exemplifies this need. This study represents the development of a robust HPLC method, engineered using analytical quality by design (AQbD), for the simultaneous quantification of UPA and its enantiomeric impurity in pharmaceutical formulations. Our AQbD approach systematically optimized chromatographic separation on a Chiralpak IG column under isocratic elution using n-hexane/ethanol mixture (70:30, v/v) at a flow rate of 1.8 mL/min, UV detection at 230 nm, and a column temperature of 40 °C. Rigorous validation using accuracy profiles confirmed the method suitability. Recognizing the growing imperative for sustainable analytical practices, we further assessed the method environmental impact through comprehensive greenness metrics, while method applicability and sustainability were assessed using Blue Applicability Grade Index (BAGI) and Red-Green-Blue 12 (RGB12) algorithms, respectively. This innovation empowers pharmaceutical manufacturers with a reliable and sustainable tool to guarantee the quality and regulatory compliance of UPA formulations, ultimately contributing to safer and more effective rheumatoid arthritis therapies.