Background: Gold is a rare and noble metal with atomic number of 79. Several techniques including both spectrometric and electrochemical methods are widely used to determine gold level in real samples. As a result of low concentration of gold in environmental samples, sample preparations such as extraction, clean-up, and pre-concentration before instrumental analysis are mandatory. Thus, a pre-treatment method (dispersive liquid-liquid microextraction (DLLME)) for gold pre-concentration and inductively coupled plasma optical emission spectrometry (ICP-OES) determination is proposed. Methods: An extraction method employing DLLME and ICP-OES has been developed for rapid separation, pre-concentration, and determination of ultra-trace amount of Au (III). The extraction of the analyte was performed in the presence of a quaternary ammonium cation, N-methyl-N,N,N-trioctylammonium chloride, (Aliquat 336) as an extractant based on ion-association extraction system.