BACKGROUND: Intravascular hemolysis with consecutive acute kidney injury (AKI) has been described after pulsed-field ablation (PFA) in atrial fibrillation (AF). OBJECTIVE: This study aimed to evaluate the risk of intravascular hemolysis and AKI after PFA with different numbers and forms of applications. METHODS: From May 2022 to August 2024, consecutive patients undergoing AF ablation were prospectively enrolled in 4 groups, including equally distributed numbers of patients undergoing PFA (pentaspline catheter) with 8 applications per vein (PFA-8), with 16 applications per vein (PFA-16), CardioFocus focal PFA, and radiofrequency ablation (RFA). Blood samples were collected immediately before and after the ablation, and on the following day to analyze hemolysis markers and indicators for AKI. RESULTS: A total of 200 patients (68.4 ± 10.8 years) could be included in the final analysis. The blood samples revealed a significant increase in total bilirubin and lactate dehydrogenase across all PFA modalities and RFA 24 hours after ablation compared with baseline levels. Potassium levels increased significantly in all PFA modalities immediately post-ablation as compared with baseline, followed by a return to approximately baseline after 24 hours. No significant potassium fluctuations were observed in RFA. Serum creatinine levels showed no significant increase in any PFA modality or RFA within the 24-hour assessment period. CONCLUSION: PFA using a multispline catheter with 8 and 16 applications per PV and focal PFA using CardioFocus (CardioFocus Inc., Massachusetts) platform showed no safety concerns with respect to hemolysis-induced AKI.