A selective separation and preconcentration method for the determination of gold ions in water and ore samples has been developed using dispersive liquid-liquid microextraction, followed by flame atomic absorption spectrometry. 4-Ethyl-1(2-(4-(4-nitrophenyl)piperazin-1-yl)acetyl)thiosemicarbazide) (NPPTSC) has been used for the first time as new chelating reagent. A mixture of ethanol (dispersive solvent) and carbon tetrachloride (extraction solvent) was used. Some parameters affecting the extraction procedure including the type and volume of the extracting and dispersive solvents, HNO3 concentration, the chelating agent amount, volume of sample, and foreign ions have optimized. Also, the complex formation between gold ions and the ligand has been investigated in a methanol-water solution (1:1) using UV-visible spectrometry. The spectrophotometric titration data showed that of Au-NPPTSC complex composition was found to be 3:2. After optimizing the instrumental and experimental parameters, we achieved a detection limit of 1.5 mu g L-1, a preconcentration factor of 50, and a linear dynamic range of 10.0-400.0 mu g L-1. The relative standard deviation obtained 2.1% at 50 mu g L-1 for gold ions (n = 10). The proposed method was successfully performed for the determination of gold in certified reference material, environmental water, and ore samples.