Background: Ergnovine (Erg) and Acetylcholine (Ach) are useful agents for assessing significant coronary artery spasm (CAS). However, comparison studies of the two agents are limited. Methods: A total of 8,093 consecutive patients (pts) who underwent Erg or Ach provocation test between November 2004 and June 2021 were enrolled. Significant CAS was defined as focal or diffuse severe transient luminal narrowing (>70%) with/without chest pain or ST-T change on ECG. We divided into two groups according to provocation agent; 1) the Erg group (n=639), 2) the Ach group (n = 7,454). We investigated whether there are differences in clinical and angiographic characteristics according to intracoronary Erg or Ach provocation test. Results: Baseline clinical characteristics were similar between the two groups except the Erg group were more elderly (58.5 years vs 56.6 years, p < 0.001) and dyslipidemia (36.0% vs 18.6%, p < 0.001) but lower rates of current smokers (16.0% vs 21.1%, p = 0.002) and alcoholics (27.1% vs 32.9%, p = 0.002). During the provocation test, the higher rates of myocardial bridge, ECG change, chest pain, atrioventricular node block, and positive CAS results but lower rates of baseline spastic change were shown in the Ach group. More pts received to lower Ach dose and higher Erg dose during the provocation test. In CAS positive patients, the Ach group had higher rates of spasm in the left anterior descending artery and mid site, but lower proximal site (table). Conclusion: Clinical and angiographic parameters were different between Erg and Ach provocation tests. Further study would be needed to validate the different parameters and different positive rates between the two agents. Table. Angiographic and Procedural Parameters during Ergonovine versus Acetylcholine Provocation Test