Tafenoquine, a synthetic analogue of primaquine, is a promising agent for the treatment of human malaria since this drug shows better antimalarial activity than primaquine in vitro and short treatment course. There are some methods for the quantification of tafenoquine in plasma, but none uses dispersive‐liquid‐liquid microextraction, an extraction technique that shows some advantages, such as miniaturization, low cost and high potential for routine application. Therefore, this study evaluated the employment of dispersive liquid‐liquid microextraction to determine tafenoquine in human plasma. The mobile phase consisted of methanol/acetonitrile/sodium acetate (10 mM, pH 6.7)/acetic acid (50:30:20:0.1, v/v/v/v) and an octadecylsilane column (15 x 4.6 mm, 5 µm) was used. The ultraviolet detection was performed at 262 nm and 1 mL/min as flow rate. The following parameters were set after method development: chloroform as extractor solvent, acetonitrile as dispersive solvent, acetonitrile:chloroform (7:3, v/v) as the final mixture, sample pH of 11.8 and extraction time of 2 min. The lower limit of quantification was 50 ng/mL and this method was linear over the range 50‐1500 ng/mL (r2 = 0.99), with satisfactory accuracy and precision, and may be useful in routine analyses.