Excess fluoride in drinking water is harmful for human health, so it is necessary to be removed. In this study the potential of a reverse osmosis (RO) membrane for defluoridation of underground water samples at different solute concentrations was studied. Because of the variation in feed water composition, its chemical properties, pH and operating conditions, effect of these parameters on the membrane separation process was studied. The results showed that pH, feed water composition, flow rate and pressure affect the membrane efficiency, and thus proper control of these factors is essential for successful operation. Optimum membrane properties and percentage rejection were determined for the RO system to minimize the overall cost of water treatment. The samples were collected from three villages in the Gurgaon district and treated at optimized conditions. The results indicate that RO membranes can be successfully used for the treatment of underground water to the desired purity level, as they remove up to 95% of fluoride present in water and also take care of other ions present in water.