In this study the effect of pH on the performance of thin film composite (TFC) nanofiltration (NF) membranes has been investigated at the relevant pH conditions, in the range of pH 1–13. TFC polyamide NF membranes have been fabricated on a polyacrylonitrile support via interfacial polymerization between piperazine in an aqueous phase and trimesoyl chloride in an organic phase. Membrane characterization has revealed that the produced membranes show a NaCl retention similar to NF-270 and Desal-5DK, a permeance in between those of NF-270 and Desal-5DK, and a slightly higher iso-electric point than NF-270 and Desal-5DK. The molecular weight cut-off of the membranes appeared to be practically constant in acidic and neutral conditions. At extremely alkaline conditions (pH > 11) an increase in molecular weight cut-off and a reduction in membrane flux has been observed. According to the Donnan steric partitioning pore model (DSPM) the change in performance in alkaline conditions originates from a larger effective average pore size and a larger effective membrane thickness as compared to the other pH conditions.