The cold-water and boiled-water soaked scrophularia soups have been prepared. The emission and excitation spectra of each scrophularia soup under different conditions have been measured at room temperature. The pH values of the different scrophularia soups have been also detected. There are obvious differences between the cold-water soaked scrophularia soup and the boiled-water soaked scrophularia. For both soups the emission wavelength increases with the wavelength of the excitation, but the peaks of the emission spectra for cold-water and boiled-water soaked scrophularia soup are different, which are 441 and 532 nm, respectively. Excitation spectrum has double peaks in the cold-water soaked scrophularia soup while only one peak with longer wavelength in the boiled-water soaked one. The pH value changes from 5.5 to 4.1. According to the organic admixture fluorescence mechanism we analyzed the reasons of the experimental results. Through heating, the interaction in different fluorescence molecular and the energy transfer process in the same fluorescence molecular become more active, and the conjugate structures and the generation of hydrogen bonds, increase. The fluorescence measurement is of value for the scrophularia pharmacology analysis and provides an analytical method for the quality identification of scrophularia soup.