A method for detecting zearalenone toxin in traditional Chinese medicines in different matrices, including extraction, purification and detection of Chinese medicine samples, said samples are extracted homogeneously at high speed with methanol-water (80:20, V/V), pH7. 0 Tween-PBS buffer solution dilution, glass fiber membrane filtration, immunoaffinity column purification. The determination method includes: high performance liquid chromatography-fluorescence detection method, high performance liquid chromatography-diode array detection method determination, spectrum and high performance liquid chromatography-mass spectrometry confirmation. Liquid chromatography conditions are: mobile phase ratio methanol: acetonitrile: water (8:46:46, V/V/V), fluorescence detector wavelength: excitation wavelength: 270nm, emission wavelength: 440nm; diode array detector wavelength: 236nm . Liquid quality conditions: Methanol-0.1% ammonium acetate water (75:25, V/V) as mobile phase, atmospheric pressure chemical ionization positive ion mode, drying gas flow rate 6.00L/min, drying gas temperature 340°C, fragmentation voltage 150V, Full scan mode scan, scan range 200-400m/z. The invention has the characteristics of accuracy, precision, reliability and the like.