A Stability-Indicating RP-HPLC Assay Method for 5-Fluorouracil

Area of Science:

• Analytical Chemistry
• Pharmaceutical Analysis

Background:

• 5-fluorouracil (5-FU) is a critical chemotherapy agent.
• Assessing 5-FU stability and purity is vital for effective cancer treatment.
• Existing analytical methods may face challenges with 5-FU degradation products.

Purpose of the Study:

• To develop and validate a robust RP-HPLC method for 5-FU determination.
• To assess 5-FU stability under various stress conditions.
• To ensure method suitability for analyzing 5-FU in bulk and pharmaceutical formulations.

Main Methods:

• Stress degradation studies (alkaline, acidic, neutral, oxidative, photolytic) were performed on 5-FU.
• Separation was achieved using a C-18 reversed-phase column with a specific mobile phase (50mM KH2PO4, pH 5.0).
• Method validation included specificity, linearity, accuracy, and precision assessments.

Main Results:

• 5-FU showed significant degradation in alkaline conditions; minimal degradation occurred under other stress conditions.
• The developed RP-HPLC method demonstrated excellent linearity (r²=0.999±0.0005) within the 10-100 µg/mL range.
• Key validation parameters, including accuracy and precision (RSD < 2.0%), confirmed method reliability, with a drug recovery of 104.69%.

Conclusions:

• The validated RP-HPLC method is highly specific and suitable for quantifying 5-FU.
• The method effectively analyzes 5-FU in the presence of its degradation products and pharmaceutical excipients.
• This analytical approach supports quality control for 5-FU bulk drug and its dosage forms.