PhD Scientific Days 2026

Pharmaceutical Sciences and Health Technologies 2.

Green Chiral HPLC Method Development for Pharmaceutical Analysis

Name of the presenter

Mhammad, Ali

Institute/workplace of the presenter

Department of Pharmaceutical Chemistry, Semmelweis University, Hőgyes E. u. 9, Budapest, 1092, Hungary

Authors

Ali mhammad¹, Gergely Dombi (Gyógyszeranalitika, HPLC)¹, Gergo Tóth¹
1: Department of Pharmaceutical Chemistry, Semmelweis University, Hőgyes E. u. 9, Budapest, 1092, Hungary

Text of the abstract

Introduction
The imperative for sustainable analytical chemistry has catalyzed innovation in green chiral high-performance liquid chromatography (HPLC) for enantiopurity determination of pharmaceuticals. This study presents a comprehensive protocol for developing eco-efficient polar organic and reversed-phase methods using polysaccharide-based chiral stationary phases and environmentally benign eluents, including dimethyl carbonate (DMC), ethanol, and water.
Aims
To devise a tiered workflow—comprising method scouting, design-of-experiments (DoE) optimization (factors: mobile phase composition, flow rate 0.5–1.0 mL/min, temperature 20–40 °C, and organic modifier), and method validation—that systematically maximizes enantioresolution (Rs), column efficiency, and sensitivity for minor enantiomer quantification (LOQ ≤ 0.02% w/w).
Methods
Representative applications included crizotinib on Cellulose-3 with ethanol:water (9:1) and thalidomide on i-Amylose-3 with DMC. All developed methods underwent ICH Q2(R1) validation, with greenness assessed via AGREE, AGSA, and mechanistic analysis of mass transfer kinetics and structural domain interactions.
Results
Crizotinib achieved Rs = 6.3, excellent linearity (r² = 0.9997), precision (RSD = 1.4%), and accuracy (98.9–101.1%); thalidomide yielded Rs = 13.8. DMC-based methods outperformed conventional acetonitrile/methanol systems (AGREE 0.77 ± 0.02 vs 0.56; Δ = 37%) and 2-propanol-based methods (AGREE 0.57), with AGSA scores 84–86. Rigid, polyfunctional analytes preferentially benefited from DMC, enabling predictive CSP–analyte–eluent matching.
Conclusion
This framework reduces solvent consumption, energy demand, and hazardous waste generation, establishing a scalable benchmark for green chiral HPLC in pharmaceutical stereocontrol and regulatory compliance.
Funding
Supported by the 2025-2.1.1-EKÖP-2025-00014 University Research Scholarship Programme of the Ministry for Culture and Innovation from the Source of the National Research, Development and Innovation Fund (G.D.). This work was funded by the National Research, Development, and Innovation Office, Hungary (grant: NKFIH FK 146930 and by the János Bolyai Research Scholarship of the Hungarian Academy of Sciences (G.T).