​As there is an increasing demand for Capsaicin from pharmaceutical firms worldwide, the aim of the present study is to find a simple, fast, and reproducible method for its extraction from the fruit of capsicum annuum. Soxhlet extraction method using n-hexan as a solvent was used. Capsaicin was isolated and purified using a combination of column chromatography (CC), and thin-layer chromatography (TLC). High-Performance Liquid Chromatography (HPLC) was used for qualitative and quantitative analysis. Capsaicin was further identified by a spectroscopic method, including nuclear magnetic resonance (NMR) and Mass spectroscopy). The extraction yield was 11.5%, and Capsaicin concentrations in the extract was 8332.3 mg/kg with purity up to 73.95%, then purification of the extract was done by column chromatography, through using two solvent system until the purified Capsaicin detected and collected. Our …​