A rapid, simple, and highly sensitive microfunnel magnetic stirring-assisted liquid-liquid microextraction coupled with HPLC was developed for determination of Riluzole (RLZ). The main extraction parameters were screened using the Plackett-Burman design and optimized by central composite design. Under optimum conditions, the calibration curve was linear over the range of 0.2-400 ng mL(-1), and RSD of 3.21% (n = 14, 20 ng mL(-1)) proved to have good reproducibility. The detection limit was 0.023 ng mL(-1) and 1.5 ng mL(-1) for aqueous and biological samples, respectively. The tap water, human plasma, and urine samples showed excellent recoveries for RLZ (97.09-101.92%) via this method.