Abstract: The present study was aimed to determine the phytochemicals of Acanthus ilicifolius by qualitative and quantitative techniques. Benzene, chloroform, acetone, ethanol and methanolic leave extract of Acanthus ilicifolius screened for their preliminary phytochemicals by standard qualitative methods. Based on this, three bioactive compounds viz., gallic acid, quercetin and lupeol were quantified using HPTLC. Results of preliminary screening expressed presence of phenols, flavonoids and triterpenoids. The Rf values of gallic acid, quercetin and lupeol are 0.21, 0.36 and 0.71, respectively. The amount of gallic acid, quercetin and lupeol was 12.26, 58.45 and 71.89 μg mL-1, respectively. In conclusion, quantification of gallic acid, quercetin and lupeol showed good resolution and separation. Further, it will be used to herbal formulation for therapeutic applications.