This study examined whether biological sex moderates the relationship between experiences of workplace culture and urinary levels of catecholamines and their metabolites. We conducted a series of regression analyses (predictors: 3-methoxytyramine (3MT), 5-hydroxyindolacetic (5HIAA), and dopamine (DA); outcomes: employee engagement and workplace culture) in a sample of 218 participants. Compared to men, women rated workplace culture less positively ( = -0.210; < 0.01) and had stronger positive associations with 3MT ( = 0.328; < 0.001), DA ( = 0.376; < 0.001), and 5HIAA ( = 0.168; < 0.01). There was a significant moderation effect between 3MT and sex on employee engagement (b = -1.76 (SE = 0.84); < 0.01), and 3MT had a positive significant association for men with engagement ( < 0.05); however, there was no significant association for women. Findings suggest that for women, less positive experiences with workplace culture could elevate 3MT, stimulating sympathetic nervous tone and potentially amplifying risks for negative health outcomes. Conversely, men who reported higher employee engagement had higher levels of 3MT, suggesting possible health risks associated with high levels of engagement, rather than lack of engagement. Overall, study findings suggested differential health risks based on biological sex, potentially impacting health risk policy development.