Plant materials and volatile extraction Habanero peppers, Red Savina, and Habanero Orange, were purchased from Shino Farm Limited Co. in 2017 3) , and all other samples were purchased from retail stores (Table 1). The samples were prepared for GC-MS analysis using a modified version of Manikharda' s method 4). The peduncle, seeds, placenta, and dissepiment were removed from each pepper, and the remaining tissues were frozen and pulverized. A portion of each prepared sample (100 g) was transferred to a 1-L beaker and slowly mixed with 1 mL 3-heptanol (100 μg/mL in methanol) , as an internal standard, 200 mL saturated aqueous NaCl (200 mL) , and 200 mL dichloromethane (DCM). The resulting mixture was gently stirred using a magnetic stirrer for 1 h at 25℃, and the DCM layer was dried over anhydrous Na 2 SO 4. Finally, volatile compounds were separated from non-volatile compounds in the DCM extract using solvent-assisted flavor Abstract: The Habanero pepper is characterized by its strong pungency and fruity aroma. The aim of the present study was to extract the volatile compounds of Habanero peppers, using solvent extraction and solvent-assisted flavor evaporation (SAFE) methods, and to analyze them using gas chromatography-mass spectrometry (GC-MS). The analysis detected 66 volatile compounds, including 6-methyl-(E)-4-heptenyl 3-methylbutanoate 1, which was reported previously, and 6-methyl-(E)-4-heptenyl 2-methylpropanoate 2, the corresponding butanoate 3, 2-methylbutanoate 4, and 6-methyl-(E)-4-heptenol 5, which were found in both Habanero and other peppers. 6-Methyl-(E)-4-heptenyl 3-methylbutanoate 1 and related compounds were synthesized. Furthermore, principal component analysis (PCA) and hierarchical cluster analysis (HCA) of the volatile profiles generally grouped the pepper samples by species and indicated that Habanero peppers are characterized by the presence of 6-methyl-(E)-4-heptenyl esters.