This study was carried out to investigate the effects of pH of sperm-washing solution, sperm-washing frequency and individual bull on concentration of hydrogen ion in sperm-washed solution and sperm acrosome reaction. The results obtained were as follow. 1. When bovine sperm was washed at 4 times with SHP solution and incubated, the difference of light absorbance between sperm-washing and sperm-washed solution(${\Delta}$Ao-${\Delta}$At) was higher in 2nd sperm-washed solution than that in the other washed solutions. 2. When sperm was thrice washed with SHP solutions of pH 5.99, 6.38, 6.78, 7.10, 7.40, 7.69, 8.15, 8.45, and 8.83, ${\Delta}$Ao-${\Delta}$At was significantly increased at pH7.69 to 8.83, and ${\Delta}$Ao-${\Delta}$At in 1st sperm washed solution was significantly higher than that in 2nd and 3rd sperm washed solution. 3. When sperm of Holstein, KNC and Hereford was thrice washed with SHP solutions of pH 5.99, 6.38, 6.78, 7.10, 7.40, 7.69, 8.15, 8.45, and 8.83, Holstein showed higher ${\Delta}$Ao-${\Delta}$At of sperm Washed solution than KNC and Hereford, and ${\Delta}$Ao-${\Delta}$At in sperm washed solution was significantly increased at 7.69 for Holstein and at 8.15 for KNC and Hereford, respectively. 4. When sperm was thrice with SHP solution of pH 6.8, 7.1 and 7.4, and then incubated in mTALP of pH 7.4 for 15 minutes, in 1st and 2nd sperm washing ${\Delta}$Ao-${\Delta}$At of sperm washed solution was significantly higher at pH 7.1 and 7.4 than at pH 6.8, and the sperm acrosome reaction of pH 6.8, 7.1 and 7.4 was 49.1, 68.8 and 72.9%, respectively. The sperm acrosome reaction of pH 7.1 and 7.4 was higher than that of pH 6.8.