Environmental Noise Directive (2002/49/EC) by the European Parliament and Council, aims to monitor and control environmental noise, by means of strategic noise maps and action plans, respectively. To that end, the number of people annoyed and sleep-disturbed are determined using harmonized noise indicator values obtained from noise maps and are reduced by executing action plans. The dose-effect relations for annoyed and sleep-disturbed people are defined by algorithms, in the Position Papers of European Commission Working Groups. In a previous study, noise maps and socio-acoustic surveys were used to form dose-effect relations for road traffic noise for Besiktas district in Istanbul, Turkey, which proved to be different from recommended relations. In this study, with the aim of analyzing the issue further, listening tests on road traffic noise annoyance were conducted for the same district. Road traffic noises were recorded according to ISO 362-1 and ISO 362-2 standards, for each vehicle type, taking into account possible vehicle speed, traffic flow type, road slope and road type typical in the area. The sound clips were formed according to the different types of traffic flow volume and traffic distribution in the area. The sound clips were then filtered with facade sound insulation values which were determined by field measurements in the area. The participants, who were all residents of the district, were asked to listen to the sound clips with headphones and to imagine they were resting inside their houses. Both verbal and numerical annoyance rating scales of ISO/TS 15666 were used for the questionnaire. The results were statistically analyzed, and responses were investigated.