A High-Performance Liquid Chromatography Method for Simultaneous Analysis of Naringin, Ascorbic Acid, and Caffeine:...

Area of Science:

• Analytical Chemistry
• Pharmacology

Background:

• Ascorbic acid, caffeine, and naringin hold therapeutic promise, particularly for colorectal cancer management.
• Existing methods often analyze these compounds individually, necessitating a more efficient approach.

Purpose of the Study:

• To develop and validate a single, robust reverse-phase HPLC method for simultaneous quantification of ascorbic acid, caffeine, and naringin.
• Ensure the method adheres to ICH Q2(R1) and USP guidelines for reliability.

Main Methods:

• Developed a reverse-phase HPLC method using a Symmetry Shield RP-18 column.
• Employed an isocratic mobile phase (75% phosphate buffer pH 3.5, 25% acetonitrile) with detection at 220 nm.
• Validated the method for specificity, linearity, precision, accuracy, and robustness.

Main Results:

• Achieved excellent linearity (R² ≥ 0.9981) and precision (RSD% < 2%) across the concentration range of 7.813–125 μg/mL.
• Demonstrated high accuracy with mean recoveries between 98%–102%.
• Method proved robust against minor variations in flow rate and pH, with system suitability parameters meeting acceptance criteria.

Conclusions:

• The developed RP-HPLC method offers a reliable, reproducible, and efficient means for simultaneous quantification of ascorbic acid, caffeine, and naringin.
• This validated method is suitable for routine quality control applications.
• The synchronized analysis supports research and development of therapeutics involving these bioactive compounds.