HSPiP, Computational Modeling, and QbD-Assisted Optimized Method Validation of 5-Fluorouracil for Transdermal...

Area of Science:

• Analytical Chemistry
• Pharmacokinetics
• Computational Chemistry

Background:

• Accurate quantification of 5-fluorouracil (5-FU) in biological matrices is crucial for pharmacokinetic studies.
• Developing efficient and robust analytical methods is essential for drug development and therapeutic drug monitoring.

Purpose of the Study:

• To establish the simplest and most efficient high-performance liquid chromatography (HPLC) method for 5-FU estimation in rat blood plasma.
• To implement Hansen solubility parameters (HSP), computational prediction, and quality by design (QbD) tools for method development and optimization.

Main Methods:

• Mobile phase selection guided by HSP predictions and experimental solubility data.
• Optimization using a central composite design (CCD) with Taguchi screening to evaluate mobile phase ratio and column temperature.
• Stability studies including freeze-thaw, short-term, and long-term stability in rat plasma.

Main Results:

• Hansen solubility parameters (HSP) and computational predictions correlated well with experimental solubility data.
• Acetonitrile (ACN) demonstrated superior suitability over methanol based on HSP and experimental findings.
• The optimized HPLC method achieved high desirability (0.978) with specific mobile phase ratio (96:4) and column temperature (40 °C).
• The method exhibited excellent robustness, reliability, and reproducibility with low limits of detection (LLOD) and quantification (LLOQ).

Conclusions:

• The developed HPLC method is robust, reliable, and reproducible for determining 5-FU in rat plasma.
• This method is suitable for pharmacokinetic parameter estimation following transdermal formulation application in rats.
• The integration of HSP and QbD significantly enhanced the efficiency and simplicity of the analytical method development.