In this work, dispersive liquid- liquid microextraction coupled with Reversed-Phase high performance liquid chromatography-diode array detector (RP-HPLC-DAD) was applied for the extraction and determination of phenylurea herbicides (PUHs) from water samples. For the extraction, 2150.0 ?L dispersive solution containing 2000 ?L dispersive solvent (acetone) and 150 ?L extraction solvent (carbon disulfide-toluene) was injected rapidly into a glass tube with conical bottom containing aqueous solution of PUHs by a 5000 ?L syringe (Gastight, Hamilton). This injection led to a cloudy water solution, caused by the fine droplets dispersion of the immiscible extraction solvent (carbon disolfide) in the aqueous sample. The result of this phenomenon was the generation of a high contact area between the aqueous phase and the extraction solvent. The final step of the microextraction procedure was centrifugation (4 min at 3500 rpm) to collect the dispersed tinny disolphid carbon droplets in the bottom of the conical test tube. After centrifugation, the sedimented phase was complectely transferred to another tube with conical bottom using a 250.0 ?L HPLC syringe. The volume of the sedimented phase was about 145.0±5 ?L. Then, the organic solvent was evaporated by a slow flow of nitrogen gas and the residue was redissolved in 25.0 ?L mixture of methanol-water (50:50) and 20.0 ?L of the solution was injected into the HPLC. The compounds of tebuthiuron, thidiazuron, fluometuron, diuron, propanil, siduron, linuron and diflubenzuron were studied. In order to obtain optimum condition of the extraction, eight parameters affecting on the extraction such as: extraction solvent type, dispersive solvent type, extraction and dispersive solvent volume, salt addition, co-solvent type, fraction of co-solvent and effect of NaCl in present of co-solvent were studied. Under optimum condition, namely, extracting solvent carbon disulfide, dispersive solvent acetone, extracting solvent volume 160.0 ?L, volume of dispersive solvent 2.00 mL, without salt addition and 30% toluene as co-solvent. The method was evaluated with respect to the LODs according to Environmental Protection Agency (EPA) Method 532. Enrichment factors and limits of detections (LODs) were ranged 11.3- 117.8 and 0.01-0.5 ?g/L, respectively. The relative standard deviations (R.S.D.s) for analysis of 5 ?g/L of each PUHs in water samples were in the range of 3/0-7.7% (n=5). Good linearity was obtained in the range of 0.2-200 ?g/L for the most compounds and correlation coffieients ranged from 0.9925-0.9995. Also, R.S.D.s for spiked water solutions at level of 5 ?g/L of each PUHs were ranged between 1.0 and 12.7% (n=5). As a result. the proposed method was used for analysis of rael samples such as tap water, well water, zayandeh rood ( Esfahan ) river water, and 3000 tonekabon river water.