In order to solve the one-sidedness and limitations of chemical purification methods, the fluorescence spectrum of Gastrodia elata powder with multiple components was theoretically analyzed and experimentally verified by combining trilinear decomposition algorithm, sum of squared error method, split-half method, and synchronous fluorescence spectroscopy. And it is demonstrated that the algorithm model for fluorescence of Gastrodia elata powder solution has three components. The results show that the first component has a characteristic fluorescence peak at excitation wavelengths of λ_ex=275 nm~280 nm, emission wavelengths of λ_em=305 nm~310 nm, and the fluorescent material corresponding to this component should be gastrodin, which has a high content in Gastrodia elata; the second component has a characteristic fluorescence peak at excitation wavelengths of λ_ex=305 nm~310 nm, emission wavelengths of λ_em=400 nm~405 nm; and the third component has a fluorescence peak at excitation wavelengths of λ_ex=355 nm~360 nm, emission wavelengths of λ_em=440 nm~445 nm; the complicated chemical separation can be replaced by the trilinear decomposition algorithm of mathematical separation, furthermore, it has the advantages of removing background noise interference, simplifying high-dimensional spectral analysis, and expressing the fluorescence characteristics of multi-component coexisting substances from an overall perspective and global features. This research provides a reference for the spectral analysis of multi-component substances.