In this study, the presence, distribution and concentration of steroidal hormones (Estrone, 17β-estradiol and 17α-ethynylestradiol) were examined in the Golden Horn Estuary (Sea, of Marmara, Turkey) for one year. The bottom and surface water samples were collected seasonally from nine stations in the Golden Horn Estuary. In order to determine the hormone levels, solid phase extraction was applied to the water samples and then these samples were analyzed by HPLC with DAD detector. Recoveries of these hormones ranged from 92-100 % for estuary water. The highest concentrations of estrone, 17βestradiol, 17αethynylestradiol were measured as 1.069 µg/L, 5.25 µg/L, 1.65 µg/L in Golden Horn, respectively. The concentrations of these three hormones are found higher in bottom water than the surface water and the highest concentrations of all hormones were determined in winter season.