The invention discloses a detection method for single-cis and double-cis lutein isomers. The method adopts a diode array detector and a YMC carotenoid C30 chromatographic column, and takes methyl alcohol, acetonitrile and dichlormethane as mobile phases, wherein the ratio of methyl alcohol to acetonitrile to dichlormethane is 50 to 30 to 20; according to the method, the time is 13 minutes, the flow velocity is 1.0mL/ minute, the column temperature is 25 DEG C, the sample injection amount is 20 microliters, the scanning wavelength is 300-500nm, and the detection wavelength is 450nm; after the method is adopted, the single-cis and double-cis lutein isomers can be well separated; furthermore, an atmospheric pressure chemical ionization (APCI) positive ion mass spectrum is utilized, the flow velocity of chromatographic column outflow components entering a mass spectrometer is 10 microliters per minute, the scanning range m/ z=80-900, the vaporization temperature is 350 DEG C, the flow velocity of dry gas is 5L/ minute, the corona current is 4 microamperes, the crushing voltage is 150 V, and the capillary voltage is 2,500V. According to the mass spectrum and optical spectrum information, the single-cis and double-cis lutein isomers are respectively determined to be 13, 15-double-cis form, 9, 15-double-cis form, 15-cis form, 13-cis form, 13'-cis form, 9, 9'-double-cis form, 9-cis form and 9'-cis form lutein. The method has the advantages of being rapid, effective, good in reproducibility, high in recycling rate and the like, and can be used for carrying out qualitative and quantitative analysis on the single-cis and double-cis lutein isomers.