HPTLC-Assisted Quantification of Active Phytoconstituents in the Aqueous Extract of Anti-hyperlipidemic Herbal Formulation

Abstract

Hyperlipidemia represents a significant metabolic abnormality characterized by elevated lipid levels that predispose individuals to cardiovascular complications. Although synthetic hypolipidemic drugs are widely prescribed, their prolonged administration is often associated with undesirable effects, encouraging the exploration of plant-based therapeutic strategies. The present study focused on the phytochemical standardization of the aqueous extract of a polyherbal formulation through quantitative estimation of selected bioactive markers using High Performance Thin Layer Chromatography (HPTLC). The formulation was prepared using Nigella sativa, Terminalia arjuna, Withania somnifera, and Linum usitatissimum, medicinal plants traditionally employed for cardiovascular and metabolic disorders. The aqueous extract was subjected to HPTLC analysis for the determination of thymoquinone, arjunolic acid, withaferin A, and linolenic acid using validated chromatographic conditions. Separation was achieved on silica gel plates with optimized solvent systems, and densitometric scanning was performed at compound-specific wavelengths. Quantitative analysis revealed the presence of thymoquinone (85.2 µg/g), arjunolic acid (12.9 µg/g), withaferin A (1.2 µg/g), and linolenic acid (0.1 µg/g) in the aqueous extract. The analytical method demonstrated satisfactory linearity, precision, and specificity, confirming its suitability for phytochemical profiling of complex herbal matrices. Peak purity and spectral overlay further ensured the authenticity of the identified constituents. The presence of these bioactive compounds supports the therapeutic relevance of the formulation and highlights its potential role in lipid regulation and cardiovascular protection. The findings establish a reliable quality control framework and provide a foundation for further experimental and clinical studies on the antihyperlipidemic efficacy of the herbal formulation.