A method for the quantitative determination of icariin, hypericin and Psoralen in mix liquor, beverage sample and tea substitutes were established by ultra-high-performance liquid chromatography tandem mass spectrometry. Samples were extracted by methanol water solution, diluted, purified, extracted, centrifuged, filtered, and determined by Thermo Acclain RSLC C₁₈ (2.6 μm, 2.1 mm×100 mm) chromatographic column. The determination was carried out under the mode of electrospray positive and negative ion multi-reaction monitoring scanning (MRM) with acetonitrile and water. The maximum linear detection concentration of 20 compounds was 500 ng/mL, the correlation coefficient (R²) was greater than 0.99, and the quantitative limit was 0.5～5.0 mg/kg. The recovery test of standard addition was carried out at three levels of 1, 2 and 10 times limit of quantitation. The average recovery rate of 20 compounds was between 72.80%～107.75%, and the relative deviation was less than 10%. Conclusion: The method is easy to operate, simple, fast, clean, high sensitivity and good repeatability.