The consumption of energy drinks has significantly increased in recent years but their adverse effects are yet to be identified. The objective of the study is to determine the acute effects of energy drink on habitual and non-habitual caffeine consumers and if caffeine consumption habit causes a difference in the hemodynamic and electrophysiological responses. We carried out a non-randomized, non-placebo controlled, three-arm parallel assignment with 48 students enrolled at Aydın Adnan Menderes University. Four main groups were determined according to the results of the application acceptance survey. Blood pressure, electrocardiogram, breath rate, haemoglobin oxygen saturation and bioelectrical impedance analysis were measured before consuming the drinks. After consuming the drinks, measurements were repeated at 30 min and 60 min. In addition, Participant's Situation Anxiety Scale was recorded. Energy drink consumption led to a significant increase in systolic blood pressure, diastolic blood pressure and heart rate for non-habitual caffeine consumers. PR interval and QRS complex were not affected significantly, however QTc interval reduced in time. Haemoglobin oxygen saturation and breathing rate did not change. Body fat percentage values of habitual energy drink consumers were smaller than the other groups. Body metabolism rate increased after energy drink consumption. There was no significant change for high-habitual caffeine consumers in all parameters evaluated. Energy drink consumption has a negative effect on hemodynamic parameters. But as the habituation level increases the impact of energy drink on hemodynamic parameters tends to decrease. Registered under ClinicalTrials.gov Identifier No: NCT04223167.