The emergence of portable global positioning system (GPS) receivers over the last 10 years has provided researchers with a means to objectively assess spatial position in free-living conditions. However, the use of GPS in free-living conditions is not without challenges and the aim of this study was to test the dynamic accuracy of a portable GPS device under real-world environmental conditions, for four modes of transport, and using three data collection intervals. We selected four routes on different bearings, passing through a variation of environmental conditions in the City of Copenhagen, Denmark, to test the dynamic accuracy of the Qstarz BT-Q1000XT GPS device. Each route consisted of a walk, bicycle, and vehicle lane in each direction. The actual width of each walking, cycling, and vehicle lane was digitized as accurately as possible using ultra-high-resolution aerial photographs as background. For each trip, we calculated the percentage that actually fell within the lane polygon, and within the 2.5, 5, and 10 m buffers respectively, as well as the mean and median error in meters. Our results showed that 49.6% of all ≈68,000 GPS points fell within 2.5 m of the expected location, 78.7% fell within 10 m and the median error was 2.9 m. The median error during walking trips was 3.9, 2.0 m for bicycle trips, 1.5 m for bus, and 0.5 m for car. The different area types showed considerable variation in the median error: 0.7 m in open areas, 2.6 m in half-open areas, and 5.2 m in urban canyons. The dynamic spatial accuracy of the tested device is not perfect, but we feel that it is within acceptable limits for larger population studies. Longer recording periods, for a larger population are likely to reduce the potentially negative effects of measurement inaccuracy. Furthermore, special care should be taken when the environment in which the study takes place could compromise the GPS signal.