Abstract: This study was conducted to formulate diclofenac sodium-loaded chitosan nanoparticles and to study the effect of formulation compositions on particle size and zeta potential of chitosan nanoparticles (CSN) containing diclofenac sodium (DC) prepared by ionotropic gelation method. It was found that the formulations containing chitosan, DC and tripolyphosphate (TPP) at a weight ratio of 4:1:1, respectively, with various pH provided various systems. At pH 5.0 and 6.0, the obtained systems were turbid because of precipitation of DC and chitosan, respectively. However, the dispersed system of CSN possessing diameter of 108±1 nm and zeta potential of 19±1 mV could be obtained at pH 5.5. These CSN also showed spherical morphology observed via a transmission scanning electron microscope. Change in weight ratio of chitosan:DC:TPP i.e. 1:1:1, 2:1:1, 3:1:1 and 4:1:1 showed that these ratios led to precipitation of particles except for the ratio of 4:1:1 providing CSN properly. The effect of Tween 80 as a stabilizer was also determined. It suggested that increment of Tween 80 concentration to 0.02% w/v could stabilize CSN at least 48 hours. However, increment of Tween 80 to 0.03% w/v led to quick precipitation of particles. The study of effect of TPP suggested that increment of TPP concentration increased particle size but decreased zeta potential. The excess TPP caused precipitation of CSN. Therefore, the optimized CSN was the CSN containing chitosan, DC and TPP at the ratio of 4:1:1and 0.02% w/v Tween 80 prepared at pH 5.5. Their particle size, zeta potential and entrapment efficiency were 128±1 nm, 15±1 mV and 45.8±2.6%, respectively.