Spiramycin removal from wastewater using four nanofiltration (NF) membranes (NF270, NF90, ESNA1-K1 and ESNA1-LF2-LD) was studied. The effects of operating pressure, feed temperature, feed concentration, cation and anion ions on the permeate flux rate and spiramycin rejection were investigated. The results show that increasing operating pressure resulted in the increase of both permeate flux and spiramycin rejection. The flux rate increased almost linearly with temperature, while the spiramycin rejection decreased. The permeate flux rate declined relatively with increasing feed concentration of spiramycin for NF270 and ESNA1-LF2-LD membranes compared with NF90 and ESNA1-K membranes. The presence of cations reduced spiramycin rejection, with the strength of influence for the NF270 NF membrane following the order Mg2+>Ca2+>K+. The presence of anions also resulted in decreased spiramycin rejection, the strength of the effect following the order NO3−>Cl−>SO42− for the NF270 membrane.