A liquid phase microextraction method-based conformation of supramolecular assembly was developed for the separation and preconcentration of trace levels of Sudan blue II. Various analytical parameters such as pH, supramolecular solvent type and volume, sample volume and matrix effect etc. were optimised. Sudan blue II concentration in the extraction phase was determined by UV-visible spectrophotometer. Under the optimised conditions, detection limit and preconcentration factor was found as 2.16 mu gL(-1) and 80, respectively. Relative standard deviation value was found 5%. The developed procedure was successfully applied for the determination of trace levels of Sudan blue II in environmental samples.