Aim: This double-blinded randomized parallel-group comparison study aimed to investigate the efficacy of an herbal desensitizing toothpaste (test group) compared to a 5% potassium nitrate toothpaste (control group) and a base toothpaste (benchmark group), with respect to dentine hypersensitivity. Materials and methods: Ninety healthy participants were arbitrarily allotted into three groups. All subjects received instructions on oral hygiene using a toothbrush with these toothpastes for a 4-week period. The subjects were evaluated at baseline, week 2, and week 4. During the visits, two hypersensitive teeth were assessed using two validated stimulus tests: a tactile test and an airblast test. Data on the percentage of positive responses to the tactile stimulus and visual analog scale (VAS) scores for air stimulation were analyzed. Results: The mean airblast VAS score and percentage of positive responses to the tactile stimulus after using the test and control toothpastes were significantly reduced compared with the benchmark. At week 4, the airblast VAS score and percentage of positive responses to the tactile stimulus decreased significantly in the test and control groups (p < 0.01), whereas the scores in the benchmark group decreased slightly. Conclusion: After 4 weeks of use, the herbal desensitizing toothpaste significantly diminished dentine hypersensitivity to the same extent as did the synthetic desensitizing toothpaste. Clinical significance: An herbal desensitizing toothpaste can reduce dentine hypersensitivity, supporting its usefulness in clinical practice.