QUALITY BY DESIGN-BASED RP-HPLC METHOD DEVELOPMENT, DEGRADATION KINETICS, AND INTEGRATED GREEN AND BLUE ASSESSMENT FOR TEPOTINIB

SYAMALA P. N. S.; SREEDEVI ADIKAY

doi:10.22159/ijap.2025v17i6.56082

Authors

SYAMALA P. N. S. Division of Pharmaceutical Chemistry, Institute of Pharmaceutical Technology, Sri Padmavati Mahila Visvavidyalayam, Tirupati, Andhra Pradesh-517502, India https://orcid.org/0009-0009-3019-9579
SREEDEVI ADIKAY Division of Pharmaceutical Chemistry, Institute of Pharmaceutical Technology, Sri Padmavati Mahila Visvavidyalayam, Tirupati, Andhra Pradesh-517502, India https://orcid.org/0000-0002-4380-2973

DOI:

https://doi.org/10.22159/ijap.2025v17i6.56082

Keywords:

Tepotinib, Reverse-phase high performance liquid chromatography (RP-HPLC), Analytical quality by design (AQbD), Degradation kinetics, Forced degradation, Greenness assessment

Abstract

Objective: Tepotinib, a recent Food and Drug Administration (FDA) approved anticancer drug, currently lacks an official pharmacopeial RP-HPLC method for quantification, and its degradation kinetics remain unexplored. This study aimed to develop a rapid, robust, and eco-friendly RP-HPLC method for the quantification of tepotinib and to investigate its degradation kinetics.

Methods: An Analytical Quality by Design (AQbD) approach was employed. The critical method parameters were screened using a Plackett–Burman design and further optimized through a Central Composite Design. Chromatographic separation was achieved on a SunFire C18 column (250 × 4.6 mm, 5 µm) using a methanol: 0.1% OPA (52:48, v/v) mobile phase at a flow rate of 1.0 mL/min with UV detection at 272 nm. Forced degradation studies were conducted under hydrolytic, oxidative, and thermal stress. The resulting degradation kinetics were then determined. The method was validated according to the ICH Q2(R2) guidelines. Greenness and sustainability were evaluated using various green analytical metrics and the Efficient-Valid-Green (EVG) framework.

Results: The developed method achieved efficient separation of tepotinib in only five minutes with a retention time of 2.4 min. Method validation confirmed excellent linearity over 22.5–157.5 µg/ml (R² = 0.9995),%RSD for intra-and inter-day precision were 0.7 and 0.5 %, mean recovery was 99.94 %. LOD and LOQ were 0.38 µg/ml and 1.14 µg/ml, respectively. Forced degradation studies revealed significant degradation under hydrolytic, oxidative, and thermal conditions, following zero-order kinetics. The greenness assessment indicated a low ecological impact, reduced solvent consumption, and minimal waste generation.

Conclusion: An AQbD-based, environmentally friendly RP-HPLC method was successfully developed and validated for quantifying tepotinib and was applied to study its degradation kinetics. This method offers environmental sustainability and is suitable for routine pharmaceutical quality control of tepotinib.

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