Objective To evaluate the effect of antihypertensive agents on primary aldosteronism (PA) screening test in the real world clinical practice; to explore the antihypertensive regimen and the corresponding aldosterone versus renin ratio (ARR) cut-off point of hypertension patients undergoing PA screening. Methods Hypertensive patients who underwent PA screening in the Department of Endocrinology & Metabolism of West China Hospital of Sichuan University between January 2010 and December 2020 were selected. According to the PA diagnostic criteria, patients were divided into the PA group and the essential hypertension (EH) group. The antihypertensive drugs, diastolic blood pressure, systolic blood pressure, blood electrolyte, plasma renin activity, plasma aldosterone, ARR were collected before and after the washout period; aldosterone before and after captopril test, and aldosterone before and after saline infusion test were collected. The above indicators of the two groups of patients were compared and analyzed. Results A total of 488 patients were included. Among them, there were 391 cases in the PA group and 97 cases in the EH group. There were statistically significant differences in age, systolic blood pressure, diastolic blood pressure, blood sodium, blood potassium, plasma renin activity, aldosterone, and ARR between the two groups (P<0.05). There was a significant difference in diagnostic results for PA screening between pre-eluting ARR and post-eluting ARR among patients taking 1-3 antihypertensive agents; however, there was no significant difference among patients taking 4 or more antihypertensive agents (P=0.547). Among the 488 patients included, calcium channel blocker and/or angiotensin converting enzyme inhibitors / angiotensin receptor interceptor were the most used antihypertensive regimen. ARR≥23.6(ng/dL)/[ng/(mL·h)] could be used as the cut-off point of ARR screening PA before elution. Conclusions When PA screening is performed in patients with hypertension, it can be directly screened for patients taking 4 or more antihypertensive drugs. As to patients taking 1-3 antihypertensive drugs, such as calcium channel blocker and/or angiotensin converting enzyme inhibitors / angiotensin receptor interceptor, ARR≥23.6 (ng/dL)/[ng/(mL·h)] can be selected as the cut-off value.