1School of Pharmacy, Suresh Gyan Vihar University, Jaipur. Email: sachinshinde18@gmail.com; ORCID: 0000-0002-4111-0439
2Associate Professor, School of Pharmacy, Suresh Gyan Vihar University, Jaipur. Email: preeti.khulbe@mygyanvihar.com; ORCID: 0009-0000-3412-6961
*Corresponding Author: Sachin S. Shinde, School of Pharmacy, Suresh Gyan Vihar University, Jaipur. Email: sachinshinde18@gmail.com; ORCID: 0000-0002-4111-0439
Analytical control of anticancer drugs requires reliable quantification methods because of their therapeutic sensitivities and clinical significance. This study applied an Analytical Quality by Design (AQbD) framework to develop and validate RP-HPLC methods for gemcitabine and methotrexate. An Analytical Target Profile was defined, and potential chromatographic factors were screened using risk assessment tools to identify the critical method parameters for each drug. These parameters were optimized using a Box–Behnken response surface design to establish a statistically supported Method Operable Design Region (MODR), ensuring robust performance under variable analytical conditions.
Gemcitabine was analyzed on an L-7 column (250 × 4.6 mm, 5 µm) with phosphate buffer (pH 6.5): methanol (97:3, v/v) at 1.0 mL·min⁻¹ in 10 min, and methotrexate on a C18 column (100 × 6 mm, 5 µm) using phosphate buffer (pH 6.0): acetonitrile (92:8, v/v) at 1.4 mL·min⁻¹ in 10 min.
The obtained MODR demonstrated wide operational flexibility and enhanced reliability for routine quality control. Validation according to ICH Q2 confirmed linearity (r² = 0.999), accuracy within 98–102%, precision with %RSD < 2%, robustness to deliberate variations, and 48-h solution stability. The developed AQbD-based methods are suitable for the routine pharmaceutical analysis of the selected anticancer drugs.
Keywords: Analytical Quality by Design, RP-HPLC, Anticancer, Method Developmen
How to cite this article: Shinde SS, Khulbe P. Analytical Quality By Design Approach in a Validated RP-HPLC Method Development for Estimation of Gemcitabine and Methotrexate in Their Dosage Forms. Int J Drug Deliv Technol. 2026;16(11s): 137-155; DOI: 10.25258/ijddt.16.11s.17
Source of support: Nil.
Conflict of interest: Nil