Abstract: This paper proposes luteolin (LUT) as a novel depressant for the flotation-based separation of scheelite and calcite in a sodium oleate (NaOL) system. The suitability of LUT as a calcite depressant is confirmed through micro-flotation testing. At pH = 9, with LUT concentration of 50 mg·L^–1 and NaOL concentration of 50 mg·L^–1, scheelite recovery reaches 80.3%. Calcite, on the other hand, exhibits a recovery rate of 17.6%, indicating a significant difference in floatability between the two minerals. Subsequently, the surface modifications of scheelite and calcite following LUT treatment are characterized using adsorption capacity testing, Zeta potential analysis, Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). The study investigates the selective depressant mechanism of LUT on calcite. Adsorption capacity testing and Zeta potential analysis demonstrate substantial absorption of LUT on the surface of calcite, impeding the further adsorption of sodium oleate, while its impact on scheelite is minimal. FT-IR and XPS analyses reveal the selective adsorption of LUT onto the surface of calcite, forming strong chemisorption bonds between the hydroxyl group and calcium ions present. AFM directly illustrates the distinct adsorption densities of LUT on the two mineral types. Consequently, LUT can effectively serve as a depressant for calcite, enabling the successful separation of scheelite and calcite.