The use of bicycles for urban trips has become increasingly important in recent years, bringing benefits such as the reduction of pollution and traffic congestion. Despite this, there are several issues that limit its establishment, such as the lack of adequate infrastructure or perceived safety. For this reason, it is important to understand the main factors which influence the use of this mode of transport and route choice. In literature, there are a number of studies that identify these factors through questionnaires, but no one analyses their choice considering the purpose of the trips for undergraduate students. To this aim, we propose a stated preferences survey to understand the most important factors influencing route choice for systematic and unsystematic travel, respectively. This survey was administered to undergraduate students of the Polytechnic University of Bari (Apulia, Italy) and results show that for home-work trips the main factor considered is travel time even with a shorter route with not good quality of infrastructure, while users of non-systematic trips tend to prefer longer routes with better characteristics. Additionally, separate cycle paths appear as determinants in both cases.