Introduction A simplified protocol for regional citrate anticoagulation (RCA) using a commercial calcium-containing replacement solution, without continuous calcium infusion, is more efficient to be used in continuous renal replacement therapy (CRRT). We aim to design a randomized clinical trial (RCT) to compare the safety and efficacy between calcium-free and calcium-containing replacement solution in CRRT with RCA. Methods Of the 64 patients receiving RCA-based post-dilution continuous venovenous hemodiafiltration (CVVHDF) enrolled from 2017-2019, 35 patients were randomized to calcium-containing group and 29 to calcium-free replacement solution group. Primary endpoint was circuit lifespan. Secondary endpoints included mortality, kidney function recovery and complications. The amount of 4% trisodium citrate solution infusion was recorded. Serum and effluent total and ionized calcium concentration were measured during CVVHDF. Results Total 149 circuits (82 in calcium-containing group and 67 in calcium-free group) and 7609 circuit hours (4335 versus 3274 hours) were included. Mean circuit lifespan was 58.1 hours (95% CI 53.8-62.4) in calcium-containing group versus 55.3 hours (95% CI 49.7-60.9, log rank P=0.89) in calcium-free group. The serum total (tCa) and ionized (iCa) calcium concentration was slightly lower in calcium-containing group during CRRT, whereas the post-filter iCa concentration was lower in calcium-free group. Moreover, mean 4% trisodium citrate solution infusion doses had no difference between groups (171.1±15.9 versus 169.0±15.1 ml/h, P=0.49). The mortality (40.0% versus 44.8%, P=0.70) and kidney function recovery rates (54.3% versus 48.3%, P=0.63) were similar in calcium-containing and calcium-free group during hospitalization, respectively. Six (3 in each group) patients showed the signs of citrate accumulation in this study. Conclusions When compared with calcium-free replacement solution, RCA based CVVHDF with calcium-containing replacement solution had similar circuit lifespan, hospital mortality and kidney outcome. Since the calcium-containing solution obviates the need for a separate venous catheter and large dose of intravenous calcium solution preparation for continuous calcium supplement, it is more convenient to be applied in RCA-CRRT practice.Trial registration Chinese Clinical Trial Registry. ChiCTR-IPR-17012629, registered on 10th September 2017. http://www.chictr.org.cn/showprojen.aspx?proj=17644