Analytical Quality by Design-Guided Optimisation and Validation of a Short-Run Stability-Indicating Reverse-Phase...

Area of Science:

• Analytical Chemistry
• Pharmaceutical Analysis

Background:

• Nonivamide (NON), a capsaicin analogue, is used in pain management.
• Limited information exists on its accurate quantification using reverse-phase liquid chromatography (RP-HPLC).

Purpose of the Study:

• To develop and validate a cost-effective, efficient, and precise RP-HPLC method for nonivamide quantification.
• To assess the greenness of the developed analytical method.

Main Methods:

• RP-HPLC method development and optimization using response surface methodology (central composite design).
• Validation according to ICH guidelines, including linearity, LOD, LOQ, precision, and accuracy.
• Greenness evaluation using the AGREE tool.
• Stress studies to assess method specificity and drug stability.

Main Results:

• Optimized conditions: 41.0% acetonitrile, 1.5 mL/min flow rate, pH 4 buffer.
• Nonivamide eluted at 6.8 min with a resolution of 13 and %RSD < 5%.
• Method validation showed linearity (2-200 μg/mL), LOD (0.67 μg/mL), LOQ (2 μg/mL), and acceptable precision/accuracy (<5%).
• Good greenness score (AGREE = 0.72).
• Nonivamide showed stability under most stress conditions, except alkaline and oxidative stress.

Conclusions:

• The developed RP-HPLC method is robust, reproducible, and reliable for accurate nonivamide quantification in pharmaceutical bulk.
• The method is environmentally considerate, aligning with green chemistry principles.
• This analytical approach supports quality control in pain management drug manufacturing.