Abstract: In order to establish method for determination of amylose contents in starch by fluorescence spectrometry. The excitation and emission spectra of amylose, and the effect of ratios of amylose to amylopectin on fluorescence intensity were determined by fluorescence spectrometry. The results showed that:The amyloseemitted fluorescence at theexcitation wavelengths of 370 nm and emission wavelengths of 422 nm. In the concentration range of 0.10~0.70 μg/mL, the fluorescence intensity of amylose presented a good linear relationship with its concentration, and the correlation coefficients was 0.9996, and the detection limit was 0.026 μg/mL. The method was applied to the determination of amylose contents in corn starchand potato with recoveries 94.6%~121.7%, and the RSD was 2.42%~3.24%. This method didn't need any biochemical reagents, and had no affected by time, temperature, coexistence of amylopectin, which was fast and simple, green and environmental friendly, low detection limit with accurate results.