The tropical donkey's ear abalone (Haliotis asinina) is routinely collected in many parts of the Indo-Pacific and has significant potential for commercial aquaculture. Although its reproductive cycle has been investigated, no studies have reported the use of chemical signals to coordinate reproduction and spawning. Using a bioassay designed with one caged individual, whilst the other is free to roam, we investigated the presence of attraction pheromones released from mature individuals. Only mature females were attracted to caged mature males with all other treatments showing no significant attraction. Those mature females also took significantly less time to reach the mature males than other pairings. Temperature influenced the response with the maximum number of positives occurring between 25 and 31°C, which coincides with the optimum physiological temperatures and the spawning times for this species. We have also shown that the attraction pheromone is present in mature male testes and induces stereotypical pre-spawning behaviours (sweeping epipodial and cephalic tentacles) as well as attracting females. Some individuals also moved towards the surface of the water, this may also be a pre-spawning behaviour. This is the first time that a sexual attraction pheromone has been reported for any haliotid species and is likely to have a critical function in the synchronization of spawning in the wild between sexes.