Aim. Based on the cortisol content in the oral fluid and the assessment of emotional stress, determine the stress level of dentists. Materials and methods. The group consisted of 25 healthy volunteers, dentists (8 men and 17 women aged 24 to 60 years) with various work experience. Participants filled in an adapted scale of the level of emotional stress before the start of the work shift. On the same day, oral fluid was collected to assess cortisol levels in the morning and evening. Results. There were no statistically significant differences in the level of emotional stress among dentists, depending on gender. The level of emotional stress in general increases with increasing work experience. With an experience of more than 20 years, emotional stress is significantly (p = 0.026) higher compared to colleagues who have been in the profession for less than 10 years, the indicator was 16.43 (±3.20) and 19.00 (±1.10) points, respectively. The level of evening cortisol in the oral fluid has a statistically significant relationship of noticeable strength with the level of emotional tension only in participants with a high level of tension (18–20 points). Conclusions. The level of emotional stress increases with increasing work experience as a dentist. To assess the level of stress, the level of evening cortisol is more indicative than in the morning, and the interpretation of the results should be carried out in conjunction with the assessment of emotional stress, as well as considering the somatic status of the subject.