UV-Spectrophotometric and RP-HPLC Method Development and Validation for Simultaneous Estimation of Gallic Acid,...

Area of Science:

• Analytical Chemistry
• Pharmacognosy
• Natural Product Chemistry

Background:

• Phytochemicals are gaining attention for their therapeutic potential and low side effect profiles.
• Gallic acid (GA), quercetin (QUE), luteolin (LUT), and apigenin (API) are key natural antioxidants found in many medicinal plants.
• Accurate quantification of these compounds is crucial for their pharmaceutical applications.

Purpose of the Study:

• To develop and validate a simple, robust analytical method for simultaneous estimation of GA, QUE, LUT, and API.
• To enable quantification of these phytoconstituents in plant extracts and pharmaceutical formulations.
• To provide reliable analytical tools for quality control and research.

Main Methods:

• Development and validation of UV spectrophotometric method for individual drug analysis.
• Development and validation of gradient reversed-phase high-performance liquid chromatography (RP-HPLC) method for simultaneous estimation.
• Validation conducted according to International Council for Harmonisation (ICH) Q2 guidelines.

Main Results:

• UV method showed good linearity and robustness for individual GA, QUE, LUT, and API.
• Gradient RP-HPLC method proved suitable for simultaneous estimation of the four phytoconstituents.
• All validation parameters met acceptable limits, though HPLC showed minor sensitivity to flow rate and wavelength variations.

Conclusions:

• Validated UV spectrophotometric and gradient RP-HPLC methods allow for simultaneous identification and quantification of GA, QUE, LUT, and API.
• These methods are applicable to crude plant extracts, pharmaceutical formulations, and biological samples.
• The developed analytical techniques support research and quality control of natural products.