Summary of - Full Factorial Design for Development and Validation of a Stability-Indicating RP-HPLC Method for the Estimation of Timolol Maleate in Surfactant-Based Elastic Nano-Vesicular Systems

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Article

Abstract

Study Background: Glaucoma is a significant cause of blindness globally, affecting over 70 million people. The disease is characterized by increased intraocular pressure (IOP), which can lead to optic neuropathy and eventual vision loss. Timolol Maleate (TM), a non-selective β-adrenergic blocker, is a first-line medication for reducing IOP by decreasing aqueous humor production. Despite its effectiveness, the delivery of TM to the inner parts of the eye remains challenging. Advances in nanotechnology-based drug delivery systems have shown promise in enhancing the ocular delivery of TM. Specifically, surfactant-based elastic vesicular systems (nanovesicles) have emerged as a potential solution, offering improved drug stability and controlled release. However, the precise quantification of TM in these systems requires a robust analytical method. Research Goals and Hypotheses: The primary goal of this study was to develop, optimize, and validate a stability-indicating Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method for the accurate quantification of TM in surfactant-based elastic vesicular systems. The research hypothesized that a novel RP-HPLC method could be developed, which would be precise, sensitive, and stable for estimating TM in the complex matrix of nanovesicles. Additionally, the study aimed to validate the developed method according to International Conference on Harmonization (ICH) guidelines, ensuring its robustness and reliability. Methodological Approach: A full factorial design using Design-Expert® software was employed to develop and optimize the RP-HPLC method. The method was developed by testing various mobile phase compositions, including different ratios of triethylamine in water (adjusted to pH 3.0) and acetonitrile. The method's robustness was assessed by varying factors such as mobile phase pH, injection volume, flow rate, and acetonitrile content. The study also included forced degradation studies to evaluate the stability of TM under various stress conditions, such as acidic, alkaline, oxidative, photolytic, and thermal environments. Validation of the method was conducted in accordance with ICH guidelines, assessing parameters like linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), and robustness. Results and Discoveries: The developed RP-HPLC method demonstrated high precision, with a linear response between 0.05 and 10 μg/mL and a correlation coefficient (R²) of 0.9993. The LOD and LOQ were found to be 0.90 ng/mL and 27.2 ng/mL, respectively. The method was specific, with a recovery rate of 89–93% for TM in both plain drug solutions and elastic nanovesicles. Forced degradation studies indicated that TM exhibited minimal degradation, except under alkaline and oxidative conditions. The total drug content in the elastic nanovesicles was estimated to be 9.53 ± 0.01 mg per 20 mL dispersion, with an entrapment efficiency of 44.52 ± 0.73%. Overall, the developed method was proven to be rapid, cost-effective, and precise for the quantification of TM in both bulk drug form and vesicular systems.

DOI: https://doi.org/10.1093/chromsci/bmab101

Publication Date: 2022

Recommended citation: Naik, S., Mullick, P., Mutalik, S. P., Hegde, A. R., Lewis, S. A., Bhat, K., ... & Mutalik, S. (2022). Full factorial design for development and validation of a stability-indicating RP-HPLC method for the estimation of timolol maleate in surfactant-based elastic Nano-vesicular systems. Journal of Chromatographic Science, 60(6), 584-594.

Publication Date

2022

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