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Objective: Seasonal variations in testosterone levels have been reported in some studies, but the results are inconsistent. In this study, we aimed to determine if clinically relevant seasonal variability in testosterone levels exists using a large cohort of men from 2 different institutions, 1 located in an area with seasons (Pittsburgh, Pa) and 1 without seasons (Miami, Fla). Methods: Using 2 institutional databases, testosterone levels were obtained for men ages 18-99 from 2010 to 2021 who had at least 2 morning testosterone levels drawn within a 2-year period. All samples were analyzed with liquid chromatography with tandem mass spectrometry. To avoid potential confounding by testosterone altering medications patients who were currently or previously on exogenous testosterone, endogenous testo stero ne-st imula ting medications, testo stero ne-su ppres sing medications, and aromatase inhibitors were excluded from the study. Results: There were 9495 and 16 171 total testosterone levels measured from Miami and Pittsburgh, respectively, with all men having 2 or more levels. There was no statistically significant variation in testosterone levels for the overall cohort in Pittsburgh or Miami, respectively. Additionally, when stratified by age group, no individual groups were found to have significant seasonal variability. Conclusion: Our findings suggest that although there is differing total testosterone levels between men who reside in 2 different climates, there is no significant variability in testosterone levels between seasons. Therefore, testosterone levels can be checked and interpreted without the need to account for the season during which they were drawn. [ABSTRACT FROM AUTHOR]