Analytical method development and validation of RP-HPLC and UPLC techniques for stability-indicating simultaneous estimation of anti-cancer combination drugs in formulated dosage forms

Abstract
The development and validation of robust analytical methods are essential for ensuring the
quality, safety, and therapeutic efficacy of anti-cancer drug formulations. With the increasing
complexity of combination therapies, reliable analytical tools are required to simultaneously
estimate multiple active pharmaceutical ingredients in formulated dosage forms. Reverse
Phase High-Performance Liquid Chromatography (RP-HPLC) and Ultra-Performance Liquid
Chromatography (UPLC) have emerged as powerful techniques that address these demands,
offering high sensitivity, reproducibility, and rapid analysis. This review consolidates recent
advancements in method development strategies, with particular emphasis on stabilityindicating assays capable of monitoring degradation products, impurities, and drug–drug
interactions. In parallel, advances in nanotechnology drug development and pharmaceutical
sciences are revolutionizing formulation design, further necessitating precise analytical tools
to evaluate nanoscale drug delivery systems and complex therapeutic
combinations.Parameters such as linearity, precision, accuracy, robustness, and specificity,
as outlined in International Council for Harmonization (ICH) guidelines, are discussed to
highlight best practices in ensuring regulatory compliance. Comparative evaluation of RPHPLC and UPLC demonstrates that while RP-HPLC remains widely adopted for routine
quality control due to its accessibility and cost-effectiveness, UPLC offers superior
resolution, faster run times, and enhanced sensitivity, making it particularly valuable for
complex formulations and stability studies. Moreover, advances in nanotechnology drug
development and pharmaceutical sciences have introduced novel delivery platforms—such
as liposomes, nanoparticles, and polymeric carriers—that demand highly sensitive analytical
validation to ensure therapeutic consistency and safety. Ultimately, RP-HPLC and UPLC
remain pivotal in ensuring the safety, efficacy, and quality of oncology therapeutics, while
future advancements in analytical sciences and nanotechnology will continue to shape the
global landscape of pharmaceutical research and quality assurance