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Background  Analysis on gender related differences in occupational stress and burnout levels usually reveal higher occupational stress and burnout levels for women compared to men, especially in male-dominated working environments. In opposition to group differentiation, more specific gender-related dimensions feminity and masculinity were used in the study to describe individual and work environment characteristics and analyze their effects.    Methods  In a cross-sectional design, survey results were linked to steroid levels in hair samples. Data was collected in a German medical services company with 146 employed women age 22–66 years ( M  = 40.48,  SD  = 10.38), 58 of them provided hair samples for steroid detection. Feminity and masculinity were measured by Gender Role Orientation Scale GTS+. Two Person-Environment fit scores in feminity and masculinity were calculated by subtracting individual from environment values. Both fit scores were proved as predictors in hierarchical linear regression models predicting burnout and work engagement as well as hair steroids cortisol, cortisone, DHEA, testosterone and progesterone detected by Liquid Chromatography-Mass Spectrometry (LC-MS/MS) as stress biomarkers. Bivariate correlations as well as moderator and mediator analysis were implemented.    Results  After considering age, role clarity, and work organization, Person-Environment fit in feminity still added significant variance explanation (β = .23,  ∆ R   2   = .05,  p  = .003) for burnout. Person-Environment fit in feminity also explained poor variance in work engagement (β = −.29,  R   2   = .09,  p <  .001). Person-Environment fit in masculinity added considerable variance explanation (β = .34,  ∆ R   2   = .12,  p  = 0.018) to cortisol levels after including quantitative demands to the model.    Conclusions  Person-Environment fit in feminity might be inspected as a predictor for burnout and work engagement. Person-Environment fit in masculinity can be taken into consideration as a predictor for hair cortisol as stress biomarker. Feminity and masculinity can be used as personality traits as well as characteristics of work environment, thus providing a particular gender-role related method of differentiation within gender groups. Also, specific methods could be derived for stress and burnout prevention and promotion of work engagement. Representative population studies with bigger samples and longitudinal surveys are needed to better explore the benefits and limitations of this approach.    Supplementary Information  The online version contains supplementary material available at 10.1186/s12995-021-00303-5.

Effects of person-environment fit of gender-role orientation on burnout, engagement and hair steroids as stress biomarkers among women