A hyphenated technique of high-performance liquid chromatography coupled with inductively coupled plasma mass spectrometry (HPLC-ICP-MS) was applied to the simultaneous determination of five organotin compounds in the sediment samples. Agilent TC-C18 column was selected; mobile phase of the HPLC was CH3CN:H2O-CH3COOH(65:23:12, V/V), 0.05% TEA, pH 3.0 at flow rate 0.6 mL min−1, and 30% O2/Ar mixed gas was added to the make-up gas, which not only reduced the carbon buildup on the cones but also protected the cones. To reduce solvent loading on the plasma, the double-pass spray-chamber was peltier cooled to −5 °C. The high RF power (1500 W) helped the samples to decompose completely. A mixed standard solution of five mixed organotin compounds from 100 μg L−1 to 1 μg L−1 was used for the method assessment. The results indicate that the correlation coefficient of calibration curves (R2) for each organotin compound was over 0.998. The international standard reference PACS-2 samples were treated by supersonic extraction with mobile phase added 0.2% tropolone. Two organotin compounds including dibutyltin (DBT) and tributyltin (TBT) were detected with this method. The values of dibutyltin (DBT) and tributyltin (TBT) were 0.99 mg kg−1 and 0.83 mg kg−1, respectively. The results showed good accordance with the certified values. The recoveries from the standard addition for five organotins were over 80%.