Objectives: Although increased reactive oxygen species (ROS) is caused by stress accelerates collagen degradation, there was no data on the relationship between stress and urinary hydroxyproline (Hyp) and proline (Pro), a good marker of collagen degradation. The purpose of this study was to evaluate the relationship between depression, anxiety, and stress (DAS) and concentrations of urinary Hyp and Pro. Methods: 97 hospital employees aged 20 to 58 were asked to fill out comprehensive self-administrated questionnaires containing information about their medical history, lifestyle, length of the work year, shit-work and DAS. Depression Anxiety Stress Scale (DASS) was applied to evaluate chronic mental disorders. Urine samples were analyzed using High Performance Liquid Chromatography (HPLC) with double derivatization for the assay of hydroxyproline and proline. Results: The mean value of Hyp and Pro concenturation in all subjects was $194.1{pm}113.4;{mu}mol/g$ and $568.2{pm}310.7;{mu}mol/g$. DASS values and urinary Pro concentrations were differentiated by sex (female > male, p < 0.05) and type of job (nurse > others, p < 0.05). In the stepwise multiple linear regressions, urinary Hyp and Pro concentrations were influenced by stress (Adjusted $r^2$ = 0.051) and anxiety and job (Adjusted $r^2$ = 0.199), respectively. Conclusions: We found that stress and anxiety were correlated with urinary Hyp and Pro concentrations. To identifying a definite correlation, further study in large populations will be needed.