A microsphere synthesized by emulsion crosslinking of polyethylenimine (PEI) was applied in the removal of ionic dyes from water. Firstly, the stability of the PEI solution-in-liquid paraffin emulsion was improved by using the mixture of Span-80/Tween-80 as the emulsifier. The optimization by 2 5-1 fractional factorial design demonstrated the most stable emulsion was prepared at 0.07 g/mL of the emulsifier dosage, 6:4 (v:v) of the oil/water ratio, 20 % (wt %) of the PEI concentration, 6:1 (w:w) of the Span-80/Tween-80 ratio with a homogenization speed of 6000 r/min for 3 min at 25 o C. Then, the polyamine microsphere (PA-M) was obtained by crosslinking the PEI emulsion with glutaraldehyde. The PA-M microsphere can adsorb xylenol orange (XO) from aqueous solution with a capacity of 1169.2±12.5 mg/g. The amount of XO adsorption remained constant in the pH range of 2.0-11.0. Adsorption kinetics analysis demonstrated that Pseudo first-order model fit the experimental data better, indicating the overall adsorption process was controlled by 'surface reaction'. The XO adsorption on the microsphere was due to an electrostatic interaction according to the results of ΔG evaluation. The value of separation factor suggested the XO adsorption was favorable. The used PA-M microsphere can be regenerated and reused in adsorption/desorption operation at least 20 times.