The current study aimed to examine the effectiveness of a web-based educational intervention based on the theory of planned behaviour (TPB) in enhancing mothers' knowledge of human papillomavirus (HPV) and its vaccine, attitudes towards the HPV vaccine, subjective norms, perceived behavioural control, and intention to vaccinate their daughters against HPV. A randomised controlled trial was conducted. The sample comprised 214 mothers of adolescents from 10 intermediate schools in Saudi Arabia. After collecting baseline data, the participants were randomly assigned to control and intervention groups. The TPB-based intervention was delivered online to the intervention group, and post-test data were collected from both groups. A mixed-effects model of covariance was used to test the effect of the intervention. There were no significant differences between the two groups at baseline. One month after delivery, the intervention was found to have a significant effect on mothers' knowledge (F (1, 211.15) = 72.40, p < .001), attitudes (F (1, 211.11) = 110.57, p < .001), subjective norms (F (1, 211) = 34.55, p < .001), perceived control behaviour (F (1, 211.12) = 144.01, p < .001), and intention to vaccinate their daughters against HPV (F (1, 211.05) = 113.64, p < .001). The study showed that the web-based intervention based on the TPB was effective in increasing mothers' knowledge, attitude, subjective norms, perceived control behaviour, and intention to vaccinate their daughters after 1 month. The findings underscore the significance of popularising and implementing such interventions to boost HPV vaccine coverage among adolescents.