Simultaneous Quantification of Caffeic Acid and p-Coumaric Acid in Typha domingensis Pers. Using a Validated High-Performance Thin-Layer Chromatography–Densitometric Method
Year: 2026
Type: preprint
Abstract: Typha domingensis Pers ., a perennial aquatic macrophyte belonging to the family Typhaceae, is widely found in wetlands and marshy habitats across tropical and subtropical regions. Different parts of the plant, including the roots, leaves, and inflorescences, have traditionally been utilized as food and in herbal medicine for treating wounds, inflammation, and bleeding-related conditions. Phytochemical investigations have revealed that the plant contains a diverse range of bioactive compounds, such as phenolic acids, flavonoids, tannins, saponins, glycosides, alkaloids, and steroids. Among these constituents, the hydroxycinnamic acids p-coumaric acid and caffeic acid are recognized as important phenolic markers because of their notable pharmacological properties. Therefore, the present study was undertaken to develop and validate a simple, accurate, precise, and robust high-performance thin-layer chromatography (HPTLC) method for the simultaneous estimation of p-coumaric acid and caffeic acid in Typha domingensis Pers. The validation of the proposed analytical method was carried out in accordance with the International Council for Harmonisation (ICH) Q2(R2) guidelines. Chromatographic separation was performed on pre-coated aluminium silica gel 60 F 254 plates using a mobile phase consisting of toluene: ethyl acetate: formic acid (7:3:0.5, v/v/v). The R f values obtained for caffeic acid and p-coumaric acid were 0.44 and 0.58, respectively, and densitometric analysis was carried out at a wavelength of 325 nm. The method exhibited excellent response over the concentration range of 200–1200 ng/band, with correlation coefficients (R²) of 0.996 for caffeic acid and 0.997 for p-coumaric acid. The detection limit (DL) was determined to be 67.01 ng/band for caffeic acid and 12.79 ng/band for p-coumaric acid, while the quantitation limit (QL) was 197.02 ng/band and 38.76 ng/band, respectively. Overall, the validated HPTLC method proved to be specific, accurate, precise, and robust for the simultaneous quantification of p-coumaric acid and caffeic acid in both their standard forms and the hydroalcoholic extract of Typha domingensis Pers.
Source: Research Square
Institution: The Charutar Vidya Mandal (CVM) University
Language: English
Subfield: Analytical Chemistry
Field: Chemistry
Domain: Physical Sciences
Open Access status: green
OpenAlex ID: W7214600562