The fluorescence spectra of ponceau 4R induced by 220-400 nm light were studied in the present paper. The result shows that ponceau 4R has four obvious fluorescence spectral peaks respectively located at 420, 530, 635 and 687 nm, each of these fluorescence spectral peaks has different best induced light, and the corresponding fluorescence spectra were listed. It was considered that this fluorescence comes from the transition n --> pi* of n electrons in the -OH and pi* --> pi of pi electrons in the naphthalene. The fluorescence spectral peaks at 420 nm come from the transition n --> pi* and the other three fluorescence spectral peaks come from pi* --> pi. But the intensity of the four fluorescence spectral peaks changes differently with the excited wavelength This paper attempted to give the expression of the four fluorescence spectral peaks based on the microcosmic mechanism. The reason for that ponceau 4R has complex fluorescence characteristic is that ponceau 4R not only has big and conjugate structure such as naphthalene and provides electron group -OH which can intensify its ability to emit fluorescence, but also absorbs electron group such as N=N which can depress its ability to emit fluorescence. Investigation on the fluorescence spectra and its characteristics will contribute to the study on the fluorescence spectra of other azo pigment and help find a new way for checking food safety.