A simple and rapid solid phase extraction procedure using a mini-column packed with modified sulfur nanoparticles as a new, efficient and reusable sorbent has been developed for the preconcentration of carbamazepine prior to its determination by high performance liquid chromatography. This method is based on the retention of carbamazepine by modified sulfur nanoparticles which are quite efficient for fast adsorption and desorption of carbamazepine. An HPLC system including C18, 250×4.6mm column, methanol-acidic water (pH=2.6 by acetic acid) (60:40) as mobile phase and UV detector (λ=276nm) was used. The effects of multiple experimental conditions such as the effect of pH, type and volume of buffer, type and volume of eluent, sample and eluent flow rate, sorbent amount and interfering ions, on the analytical performance of the method were investigated. The calibration curve was linear in the range of 0.5-200ngmL-1 and LOD of the proposed method was found to be 0.16ngmL-1. The procedure was successfully applied for the determination of carbamazepine in pharmaceutical samples, human plasma and breast milk.