Noise is a common problem in hospitals, and it is known that social behavior can influence sound levels. The aim of this naturally-occurring field experiment was to assess the influence of a non-talking rule on the actual sound level and perception of patients in an outpatient infusion center. In a quasi-randomized trial two conditions were compared in real life. In the control condition, patients (n = 137) were allowed to talk to fellow patients and visitors during the treatment. In the intervention condition patients (n = 126) were requested not to talk to fellow patients and visitors during their treatment. This study measured the actual sound levels in dB(A) as well as patients’ preferences regarding sound and their perceptions of the physical environment, anxiety, and quality of health care. A linear-mixed-model showed a statistically significant, but rather small reduction of the non-talking rule on the actual sound level with an average of 1.1 dB(A). Half of the patients preferred a talking condition (57%), around one-third of the patients had no preference (36%), and 7% of the patients preferred a non-talking condition. Our results suggest that patients who preferred non-talking, perceived the environment more negatively compared to the majority of patients and perceived higher levels of anxiety. Results showed no significant effect of the experimental conditions on patient perceptions. In conclusion, a non-talking rule of conduct only minimally reduced the actual sound level and did not influence the perception of patients.