This application demonstrated the automated and fast method development capability of the ACQUITY UPLC with 2D Technology for the analysis of synthetic cannabinoids in urine, plasma and edible samples. The quantification limit was set at 50 ppt using a 1.0 g of sample. The micro extraction protocol offered the option to evaluate several elution parameters in a short time period. The elution optimization was completed within a 4 hrs hands-on work and the 2D LC results were analyzed using an over-night run multi-methods sample list (18 hrs). With the extraction protocol optimized, the final protocol produced a clean extract in 30 minutes without any evaporation to dryness and reconstitution into initial mobile phase conditions.