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The TFC RO/NF membranes are the predominant membranes applied in desalination, and heat curing is an essential step in its industrial production, which increases the polyamide cross-linking degree and facilitate packing but leads permeability decrease. Nevertheless, the mechanism of heat curing on TFC membranes permeation resistance changes is still ambiguous. Figuring it out will aid in crafting a more effective optimization strategy in high flux membrane preparation. Herein, we prepared a range of TFC RO/NF membranes with various monomer concentrations on different substrates and quantified the permeation resistance changes by a resistance-in-series model. For nascent TFC RO/NF membranes, the PA layer provided almost all permeation resistance. After heat curing, the substrate exhibited a marked increase in permeation resistance. The change in the permeation resistance composition was further exacerbated by the decrease in monomer concentration. For TFC NF membranes with heat curing, the proportion of substrate permeation resistance increased significantly, rising from 31.62 % to 75.05 %. This study made a comprehensive analysis of the impact of the permeation resistance in TFC RO/NF membranes caused by heat curing, enriched the insight into the impact of heat curing on TFC membranes, and helped to prepare TFC membranes with better desalination performance. [Display omitted] • The resistance proportions of RO/NF membranes are quantitatively revealed. • The substrate resistance increases significantly due to heat curing. • Low monomer concentration results in high substrate resistance. [ABSTRACT FROM AUTHOR]