The invention relates to an eye drop used for releasing eye fatigue and a preparation method thereof. The eye drop comprises following raw materials: radix rehmanniae, rhizoma atractylodis, crab grass, serissa japonica, jasmine, red-spotted stonecrop, wolfberry, astragalus smicus, python gall, cocklebur, chamomile, concha haliotidis, Taiwan beautyberry, globe amaranth, pearl and weathered sodium sulfate powder. The eye drop is a pure traditional Chinese medicine preparation; is prepared by modern advanced pharmaceutical technologies; has efficacies of clearing away heat and purging pathogenic fire, relieving spasm and removing eyewinker, and nourishing the liver and improving visual acuity; and can be used for treating diminution of vision and pseudomyopia of teenagers, and reliving visual fatigue of eyes, and symptoms caused by visual fatigue such as conjunctival injection and eye itch. Compatibility of the selected raw materials is appropriate, curative effects are significant, safety performance is excellent, no side effects exist, cost is low, and the eye drop is worth of wide popularization and application.