Developing new, eco-friendly solvents which would meet technological and economic demands is perhaps the most popular aspects of Green Chemistry. Natural deep eutectic solvents (NADES) fully represent green chemistry principles. In this sense a new green vortex-assisted NADES-based microextraction (VA-NADES-ME) was established, for the first time, for the extraction of lamotrigine from human plasma samples using NADES, as a green extractant alternative to hazard organic solvents. Then extract was determined by high performance liquid chromatography with ultraviolet detection (HPLC–UV). The mixture of (choline chloride/ethylene glycol) as NADES was found to be the best candidate for extracting lamotrigine. Grey relational analysis-based Taguchi method was utilized as a chemometric protocol for multiobjective optimization of different HPLC performance parameters. The optimum mobile phase was a mixture of water with 0.3 % triethylamine in 0.02 M H3PO4 (adjusted to pH 5.5) and acetonitrile (70:30 v/v) with an overall pH 6. Finally, the confirmation test was performed to validate the predicted optimal results and established an improvement of 32.84 % from the initial design. The mobile phase was eluted in an isocratic mode with a flow rate of 0.8 mL/min and UV detection at 310 nm. The retention times of lamotrigine and tinidazole (internal standard) were 4.7 and 3.4 min, respectively. Proposed method was linear over the concentration range of 0.1–15.0 μg mL−1. The greenness profile was assessed using Analytical GREEnness metric approach which assured the greenness of the proposed method compared with other reported methods.