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Currently, most nanofiltration membranes exhibit satisfactory separation performance on the anions or cations, and few membranes could concurrently separate anion and cation. Therefore, preparing nanofiltration membranes with excellent separation performance for both anionic and cationic ions remains a challenge. Herein, we have developed a series of nanofiltration membranes using conventional interfacial polymerization with different ratios of trimesoyl chloride (TMC) and piperazine (PIP) concentrations. In the interface polymerization process, the amine groups carried by PIP imparted a positive charge, while the hydrolysis of acyl chloride groups resulted in the generation of carboxylic acid groups that introduced a negative charge. The mix-charged nanofiltration membrane could be fabricated by varying PIP/TMC concentration ratios. The optimized membrane exhibited high rejection rates for bivalent ions (Na 2 SO 4 rejection of 96.6 %, MgCl 2 rejection of 93.0 %), and its selectivity for Mg2+/Li+ mixed solution was improved significantly. This work provides a novel approach for the development of nanofiltration membranes capable of effectively separating mono-/divalent ions. [Display omitted] • Fabrication of mixed-charged polyamide membranes via regulating the PIP/TMC ratio directly. • High PIP/TMC ratio leads more unreacted amine groups in the polyamide which was beneficial to MgCl 2 rejection. • Such NF membrane showed excellent separation efficiency to Mg2+/Li+ and SO 4 2−/Cl−. [ABSTRACT FROM AUTHOR]