Background : Coffee is one of the favorite brewed drink in the world where is distributed in Latin America, Southeast Asia, Southern Asia and Africa. Coffee has an effective antioxidant ability and reported about that. In this study, it was analyzed by using high performance liquid chromatography (HPLC) to establish the method about content of caffeine, chlorogenic acid, caffeic acid and p-coumaric acid in coffee.
Methods and Results : Coffee was extracted with 70% EtOH in room temperature and evaporated at 45℃. All standard and sample extract were melted and diluted with 15% MeOH. Mobile phase was prepared using water with 0.01% phosphoric acid and MeOH. All standard and sample were analyzed with gradient elution (0 min : 15% MeOH, 35 min : 30% MeOH). The chromatograms were monitored at 272 and 320 ㎚. HPLC reported linear equation that based on the calibration curve for each standard compound (caffeine : Y = 1.04e + 004X – 3.21e + 003, R2 = 0.999890. chlorogenic acid : Y = 2.86e + 004X – 8.24e + 003, R2 = 0.999891. caffeic acid : Y = 2.07e + 004X – 1.21e + 004, R2 = 0.999894. p-coumaric acid : Y = 3.24e + 004X – 1.10e + 004, R2 = 0.999897). Standard compounds were determined with qualitative and quantitative analysis. The retention time of each peak of standard compounds were separated by chromatogram.
Conclusion : In this study, we determined that the analysis method of compounds in coffee. In addition, we have confirmed that separation about the retention time of each peak of caffeine and chlorogenic acid in different solvent condition depending on acid buffer. This method can be use to determine standard compound in coffee.