The increasing number of cyclists in cities around the world results in a greater focus on bicycle traffic. Next to traffic volume, the main characteristic of traffic used in road safety analysis, infrastructure planning, design, etc. is its speed. Bicycle speed is strongly affected by the type of bicycle facility, motor vehicle traffic parameters (volume, speed, share of heavy vehicles), trip motivation, weather conditions, etc., and therefore it is difficult to estimate. Traditionally, bicycle speed is determined directly using speed radar or indirectly, as a quotient of measurement base length and travel time calculated using a stopwatch or video technique. There are also researches where bicycle speed was estimated based on GPS sources, mainly mobile apps. However, depending on the GPS source and the group of cyclists, bicycle speed gained from GPS data can be different from the speed of regular cyclists (due to different levels of experience or types of bicycle). In the paper, the relationships between bicycle speed obtained from empirical measurements and two different GPS sources, which were bikesharing system (Wavelo) and Strava app, were analysed. In total 18 research sites were selected different in terms of bicycle facility (bicycle path, shared pedestrian/bicycle path, contraflow lane) and element of road network (road segment, bicycle crossing with or without traffic signals). Two-tailed test for two means was conducted to analyse the statistical significance of differences in bicycle speed estimated based on GPS data and empirical measurements using video technique. It showed that Wavelo and Strava speeds are by 17.4% lower are by 23.1% higher than the speeds of regular cyclists respectively. Two linear regression models describing relationships between bicycle speeds from empirical measurements and GPS data were developed. The results show that the variance of bicycle speed is almost 80% described by the variance of Wavelo speed and 60% described by the variance of Strava speed, which suggests that bicycle free-flow speed can be estimated based on GPS data either from bikeshare system or dedicated app.