This research was aimed at analyzing the acute effects of Arabica black coffee consumption on blood glucose, insulin, and serum cortisol levels, as well as determining the pharmacological effects of black coffee as an antihyperglycemic. A randomized control trial with healthy female subjects was used in this study. There were 20 volunteers in total: 9 as the control group and 11 as the trial group. The treatment included brewing 10 grams of Gayo Arabica black coffee powder with 150 ml of boiling water. Blood glucose, insulin, and cortisol levels were measured twice, before and after 60 minutes of coffee consumption. An independent sample t-test (p < 0.05), Pearson correlation test (p < 0.05), and simple linear correlation test (p < 0.05) were used to analyze the data. Blood glucose levels and serum cortisol levels decreased significantly after coffee consumption in the trial group (p = 0.002* and p = 0.001*). There was no significant negative correlation between glucose and insulin levels (r = -0.122; p = 0.721). On the other hand, there was a significant positive correlation between cortisol levels and blood glucose (r = 0.651; p = 0.002*). In conclusion, a single cup of Gayo Arabica black coffee reduces blood sugar and serum cortisol levels, but does not increase serum insulin levels. Blood glucose levels correlate positively with serum cortisol levels in healthy female.