High-Performance Thin Layer Chromatographic (HPTLC) Quantification of Gallic Acid from Emblica officinalis Fruit Extract

Abstract

A dependable analytical method for phytochemical standardization and quantitative marker compound quantification in herbal formulations is High-Performance Thin Layer Chromatography (HPTLC). Emblica officinalis (Amla) is a therapeutically important medicinal fruit, widely used in Ayurvedic and nutraceutical preparations, with gallic acid recognized as its major bioactive constituent contributing to antioxidant potential. The present study aimed to develop and validate a simple, selective, and precise HPTLC method for the quantification of gallic acid in the methanolic fruit extract of E. officinalis. Methanolic extraction of dried fruit powder yielded 6.32% extract using a standard gravimetric method. Chromatographic separation was achieved on silica gel 60 F₂₅₄ plates using toluene: ethyl acetate: formic acid (5:4:1, v/v/v) as the mobile phase. A sharp and well-resolved peak of gallic acid was observed at Rf = 0.43 ± 0.02, matching the standard marker. Quantification was performed through densitometric scanning at 254 nm, and the calibration curve displayed excellent linearity with a correlation coefficient (R² = 0.998) over the selected concentration range. Validation of the method in accordance with ICH Q2(R1) guidelines confirmed high precision, accuracy, sensitivity, and reproducibility. The results indicate that gallic acid is a major phenolic constituent in E. officinalis fruit extract and support the utility of the developed HPTLC protocol for routine quality assessment and standardization of herbal raw materials and formulations containing E. officinalis.