The prevalence of dentin hypersensitivity in Taiwan has been reported up to 38%. The aim of this study was to evaluate the clinical efficacy of desensitizing toothpaste containing 8.0% arginine and calcium carbonate in treating dentin hypersensitivity. Forty-three participants with established two hypersensitive teeth were enrolled with informed consent. Air blast sensitivity assessments were conducted at baseline and again after four and eight weeks of twice-daily product use. The use of desensitizing toothpaste was found to significantly reduce dentin hypersensitivity (P < 0.001). The one-way analysis of variance analysis followed by Tukey's test indicated that this desensitizing toothpaste had a time-to-improvement distribution (P < 0.05). Taken together, the toothpaste containing 8.0% arginine and calcium carbonate provides a significant reduction in dentin hypersensitivity.