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Emotional stress negatively affects the quality of a person's daily life. From a physiological point of view, stress is expressed in the excitation of the hypothalamic-pituitary-adrenal cortex axis, which leads to the release of the hormone cortisol into the blood. We developed a lateral flow immunoassay to detect cortisol in human salivary fluid and tested it on 10 healthy volunteers daily for about one month (n = 293 saliva samples). Cortisol was detected in concentrations ranging from 1 to 70 ng/mL. Salivary cortisol levels were confir