RP-HPLC Method Development and Validation for Simultaneous Quantification of Ascorbic and Gallic Acids in Natural Extract-Based Granules

Background

Ascorbic acid and gallic acid are known for immune-boosting and antioxidant properties.

Objectives

To develop a straightforward, sensitive, validated, and economical reverse phase liquid chromatographic technique for the simultaneous measurement of ascorbic and gallic acid in granule formulation and natural extracts (Amla and Chyavanprash). These compounds are essential for their therapeutic benefits.

Materials and Methods

Ascorbic and Gallic acid were separated from the complex mixture of various polyphenols present in natural extract. At 30°C, these two compounds were separated using a Thermo C18 analytical column with an isocratic program consisting of two solvents-A & B. The ratios of Solvent- A (0.3% orthophosphoric acid in HPLC grade water) and Solvent-B (80% acetonitrile in HPLC grade water) were 85:15 v/v, and the flow rate is 0.5 mL/min. using a UV-PDA detector at a single wavelength of 250 nm.

Results

In accordance with ICH guidelines, the developed method was validated for linearity, precision, accuracy, robustness, Limit of Detection (LOD), & Limit of Quantification (LOQ). The granule formulation has 0.261% gallic acid and 0.079% ascorbic acid.

Conclusion

Ascorbic acid and gallic acid in natural extracts and granule samples were successfully quantified and validated using the RP-HPLC method. The method proved it’s simple, precise, sensitive, rapid and robust.