A method based on the TMS derivatives and acidic hydrolysis was developed for the simultaneous determination of free and conjugated steroidal hormones in surface water. A silylation of five natural and two synthetic steroidal hormones was achieved with N-methyl-N-(trimethylsilyl) trifluoroacetamide/$NH_4I$ (1000:3) under catalysis of dithioerythritol for 60 min at $80^{circ}C$. TMS derivatives of the steroid hormones containing multifunctional groups offer a single derivative product under this condition. The accuracy of the analytes was in the range of 87 to 110% at a concentration of 20 and 50 ng/L with relative standard deviations of less than 10%. The method detection limit was in the range of 0.01 to 0.02 ng/L for surface water. Natural steroidal hormones were detected in a concentration range of 0 to 1.03 ng/L in free form and 0 to 14.6 ng/L in conjugated form, respectively. We found that most of the natural hormonal steroids exist in conjugate forms (43 to 100%) in river water.