A simple，rapid and effective method for the quantitative analysis of quercetin in Gynostemma pentaphyllum was established by using excitation-emission matrix (EEM) fluorescence coupled with second-order calibration based on Parallel factor analysis(PARAFAC) algorithm，alternating trilinear decomposition (ATLD) algorithm and self weighted alternating trilinear decomposition (SWATLD) algorithm.Al(Ⅲ) was used as fluorescence sensitizer to increase the quercetin fluorescence intensity.The core-consistency diagnostic(CORCONDIA) test indicated the number of components should be two，one of the components corresponds to the quercetin，and the other is related to a coexistence component of traditional Chinese medicine.The content of quercetin by PARAFAC，ATLD，SWATLD with component number N=2 are 0.274 9%,0.272 9% and 0.272 7%，and the average recoveries were 96.3%，103.1% and 96.8%，respectively.The resolved quercetin spectra overlap almost completely with real spectra.The experimental results showed that the chemometrics “second order correction” method was simple，rapid，high efficient and low cost，and could be used for complex component qualitative and quantitative analysis.In addition，the influence of the inner filter effect on fluorescence intensity was discussed，and the results revealed that ultraviolet correction could effectively improve the accuracy of quantitative analysis.