In the present study, the N, O and S donor Isatin-3-(4-phenyl-3-thiosemicarbazone) (IPTS) formed hydrophobic complexes with Pd(II) and Co(II). Characterization of these complexes was carried out using various instrumental performances such as elemental analysis, Fourier transform infrared spectroscopy (FTIR), and UV-vis spectrophotometry. A micelle-mediated extraction (MME) method developed for the separation and enrichment of trace levels of palladium and cobalt was combined with inductively coupled plasma optical emission spectroscopy (ICP OES) to measure trace levels of palladium and cobalt in real samples. Hydrophobic Pd(II)-IPTS and Co(II)-IPTS complexes are extracted into the Triton X-114 rich phase. Parameters affecting both complexation and extraction, such as sample pH, IPTS and Triton X-114 concentrations, equilibration temperature, and time, are optimized. After the separation process, the Triton X-114 rich phase was diluted with acidic ethanol. Analyte concentrations were then determined by ICP OES. Under the optimal conditions chosen, the LOD, and LOQ for Pd(II) were 54 ngL-1, and 164 ngL-1, respectively. The LOD and the LOQ for Co(II) were 143ngL-1 and 432 ngL-1, respectively. This MME method was applied to real samples and recoveries of more than 95% were obtained in the spiked samples with a preconcentration factor of 100 and relative standard deviation less than 5% (n=5). The plausible mechanism of interaction between Triton X-114 and Pd(II)-IPTS and Co(II)-IPTS complexes is elucidated