Objective: Development of micelle mediated cloud point extraction (CPE) and preconcentration of aurintricarboxylic acid (ATCA) (a viral inhibitor). Methods: The method is based on the ion-pair formation between ATCA and a cationic surfactant tetrabutylammonium bromide (TBAB), which is extracted into the Triton X-114 (TX-114, a non-ionic surfactant) at a pH 4.4. The effect of different important parameters like pH, the concentration of surfactants (TX-114 and TBAB), salt, temperature, and time for the extraction of ATCA was optimized. Results: Extraction efficiency of 90.28 % for CPE of ATCA was obtained using mixed micelles of TX-114 and TBAB. The linear range and limits of detection for ATCA was found to be 8.44-84.47 µg ml-1 and 8.14 ng ml-1, respectively. Conclusion: The suggested CPE method has been applied to the determination of ATCA in aqueous solutions.