Arisaema tortuosum (Wall.) Schott holds significant ethnomedicinal value, yet its phytochemical profile has remained scientifically underexplored. This study provides a comprehensive analysis of its tuber extracts, revealing substantial solvent-dependent variations in composition. Extraction through maceration yielded 2.5% (chloroform), 2.9% (ethyl acetate), 6.3% (ethanol), and 11.2% (aqueous) w/w of crude extract. Phytochemical screening demonstrated the presence of alkaloids, flavonoids, phenols, and tannins across various extracts. Quantitative analysis established the ethanol extract as superior, containing the highest total phenolic (0.878 mg GAE/100 mg) and flavonoid (0.805 mg QE/100 mg) content.

A novel HPTLC method for lupeol quantification was developed and validated per ICH guidelines, showing excellent linearity (R² = 0.999) across 50-175 ng/spot. The method demonstrated high precision (intra-day %RSD 0.04-0.09, inter-day %RSD 1.60-1.75) and accuracy (recovery 99.92-100.05%), with LOD and LOQ of 1.22 and 3.70 ng/spot respectively. Lupeol was identified at Rf 0.67 and quantified highest in ethanol extract (0.067% w/w), followed by chloroform (0.055%) and ethyl acetate (0.052%) extracts. GCMS analysis confirmed lupeol identity through matching retention times (sample: 31.85 min, standard: 31.86 min) and characteristic molecular ion (m/z 426).

This study validates A. tortuosum as a rich source of bioactive compounds. The ethanol extract emerges as a particularly promising candidate for further drug development. These findings successfully bridge traditional knowledge with scientific validation, advocating for deeper exploration of its phytochemical mechanisms. The proposed HPTLC method provides a dependable tool for quality control and standardization of this vital medicinal plant.