PhD Scientific Days 2025

Pharmaceutical Sciences and Health Technologies I.

Green HPLC Strategy for Chiral Resolution of Crizotinib on Polysaccharide-Based Columns

Name of the presenter

Mhammad Ali

Institute/workplace of the presenter

Department of Pharmaceutical Chemistry, Semmelweis University

Authors

Ali Mhammad^1,2, Gergely Dombi^1,2, Máté Dobó^1,2, Zoltán-István Szabó^3,4, Gergő Tóth^1,2

1: Department of Pharmaceutical Chemistry, Semmelweis University, Hőgyes E. u. 9, Budapest, 1092, Hungary
2: Center for Pharmacology and Drug Research & Development, Semmelweis University,Budapest, Hungary
3: George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Târgu Mureş, Romania
4: Sz-imfidum Ltd, Lunga, 525401, Romania

Text of the abstract

Introduction: With the growing emphasis on environmentally friendly analytical
practices, green approaches to chiral separation—particularly high-performance liquid chromatography (HPLC) —have gained increasing attention as essential tools for determining the enantiomeric composition of chiral pharmaceutical compounds.

Aims: To develop a green and efficient HPLC method for the enantioseparation of
crizotinib, a tyrosine kinase inhibitor targeting anaplastic lymphoma kinase.

Methods: The enantioselective performance of seven polysaccharide-based chiral
stationary phases was evaluated, including three amylose-derived (Lux Amylose-1, Lux
i-Amylose-1, Lux Amylose-2) and four cellulose-derived columns (Lux Cellulose-1, Lux
Cellulose-2, Lux Cellulose-3, and Lux Cellulose-4), under both polar organic and
reversed-phase modes.

Results: Among these, Lux Cellulose-3, which incorporates cellulose tris(4-methylbenzoate) as the chiral selector, demonstrated the best resolution using an
ethanol–water mixture with 0.1% diethylamine as the mobile phase. Chromatographic
parameters (mobile phase composition, flow rate, and temperature) were optimized using a full factorial screening design. Under optimal conditions (Lux Cellulose-3 column, ethanol/water/diethylamine 92/8/0.1 v/v/v, 0.5 mL/min flow rate at 40 °C column temperature), baseline enantioseparation was achieved. The method was validated in accordance with current guidelines. The greenness of the proposed method was confirmed through Analytical Eco-Scale, Complex GAPI, and AGREE metrics, showing superior environmental performance compared to a previously published method.

Conclusion: These findings highlight the method's reduced solvent hazards and waste generation, supporting its classification as a green analytical procedure.

Fundings: This work was funded by the National Research, Development, and Innovation Office, Hungary (grant: NKFIH FK 146930).