Raman spectroscopy is a valuable method to explore the structural characteristics of molecular and crystal, which has been widely used in food safety, petrochemical and other fields due to its advantages of fast, non-destructive, small sample consumption, no pretreatment and adaptability. However, it is often interfered with by the fluorescence background when applied, which leads to the decrease of the Raman signal. The Raman signal would be submerged in the fluorescence background in the worst situation. In order to solve the problem that the fluorescence background interference in the application of Raman detection, this research have designed the optical path coupling design of multi-wavelength Raman spectrum with dichroic mirror in the instrument and developed a multi-wavelength fluorescence elimination Raman spectrum detection system combining near Red Raman spectrum and frequency shift differential Raman spectroscopy. The near-infrared Raman spectrum is designed with a 1 064 nm laser source, and the frequency shift differential Raman spectrum is composed of 784.5 and 785.5 nm laser sources. By comparing the intensity and peak stability of synchronous test and successive time-sharing test, the synchronous testing performance of multi-wavelength de fluorescence Raman spectrometer was verified; The single wavelength Raman, near-infrared Raman and frequency shift differential Raman spectra, were compared and analyzed. For acetone, acetonitrile and other samples with weak fluorescence background, single wavelength Raman spectroscopy can be used for the quantitative and qualitative analysis; Near-infrared Raman spectroscopy can be used for the quantitative and qualitative analysis of samples with similar fluorescence background and Raman signal intensity such as edible oil and red plastic particles; For the samples with strong fluorescence background such as red wine and brown plastic particles, the samples can be qualitatively analyzed by near-infrared Raman spectroscopy and differential Raman spectroscopy. The results showed that based on conventional single wavelength Raman spectroscopy technology, we combined the two fluorescence interference suppression technologies through the development of multi-wavelength de fluorescence Raman spectroscopy detection system. This research will expand the range of the application field and sample detection.