An increasing interest in the research and isolation of different flavonoids from plants due to their beneficial and potential therapeutic effects as reported in many literatures. Achillia santolina belongs to the family Astaraceae, is a known plants rich in flavonoids and other phytochemicals that exhibit many biological activities like cytotoxicity, anti-inflammatory, antimicrobial, cholesterol level reduction and anticancer activity. The plant total flavonoids had been extracted by reflux condenser apparatus with 10% aqueous HCl then aglycone moiety was obtained with ethyl acetate. Quantitatively, the extraction yield of ethyl acetate (EA) fraction was 0.7689gm/40 gm (1.922%) of dry plant materials.While the total flavonoid content was (22.01362±2.1mg/g) of Quercetin equivalent of dry extract. For quantitative estimation of individual aglycons, Luteolin showed the highest concentration in EA fraction followed by Apigenin, Isorhamnetin, Quercetin, and Kaempferol with concentrations of (0.223±0.17), (0.184±0.18), (0.151±0.14), (0.148±0.08), and (0.024±0.10) μg/mg of dry extract, respectively. The current study showed Achillia santolina grown in Iraq is a valuable plant and considered as a rich source of flavonoids for combating many diseases.