Background: Phlebotomy is an essential initial examination needs to be done correctly to diagnose and treat the patient. Venipuncture is an example of invasive medical procedure categorized as the most uncomfortable one. The most dominating populations in Indonesia are productive age group, aged 18-65 years old. Methods: Quasi-experimental was used in this study. Populations in this study were median cubital venipuncture patients in RS PKU Muhammadiyah Yogyakarta aged 18 until 65 years old, of 36 respondents divided into 2 groups of 18 patients in treatment group and 18 patients in control group. Patients filled informed consent before the treatments were done. In treatment group, ethyl chloride spray was given before venipuncture. Pulses and VAS scores were noted before and during venipuncture. Results: Mean of VAS score showed higher score in control group 3,67 in score compared with treatment group in 0,06 in score. Mean subtractions of pulses in control group were higher than treatment group, 9,6667 compared to 2,333. Based on statistic test in both variables, Asymp. Sig (2-tailed) of 0,000 (<0,05) was obtained, statistically significant in meaning. Conclusion: Administration of ethyl chloride spray was effective towards pain level reduction of median cubital venipuncture.