Objectives: To examine the relationship between levels of testosterone, free testosterone, and sex hormone binding globulin (SHBG) in women diagnosed with polycystic ovary syndrome (PCOS) and a control group. Study Design: Observational study. Participants and methods: The study included 50 women diagnosed with PCOS and 50 women without a PCOS diagnosis, aged between 18 and 44 years. It was conducted by searching Laboratory Information System (LIS) and Hospital Information System (HIS) databases from 2021 to 2023. All significant data, including concentrations of parametres and the presence of certain symptoms, were recorded in an Excel table. Results: Elevated levels of testosterone and free testosterone, along with decreased levels of SHBG, were recorded in women with PCOS, and these parameter levels differed from those in women without PCOS. Within the PCOS group, it was observed that patients with obesity and insulin resistance (IR) had elevated levels of free testosterone and decreased levels of SHBG. Patients in the PCOS group, both with and without hirsutism showed no differences in these parameters. Conclusion: Women with PCOS have elevated levels of testosterone and free testosterone, along with decreased levels of SHBG, which significantly differ from those in women without PCOS. No statistically significant association was found between these parameters and hirsutism, but a connection between elevated free testosterone, reduced SHBG levels, and obesity was established. Free testosterone and SHBG may serve as useful biomarkers for diagnosing IR in PCOS.