A second-order multivariate calibration approach, based on a combination of unfolded-partial least-squares with residual bilinearization (U-PLS/RBL), has been applied to fluorescence excitation-emission matrix data for multicomponent mixtures showing inner filter effects. The employed chemometric algorithm is the most successful one regarding the prediction of analyte concentrations when significant inner filter effects occur, even in the presence of unexpected sample components, which require strict adherence to the second-order advantage. Results for simulated fluorescence excitation-emission data are described, in comparison with the classical approach based on parallel factor analysis and other second-order algorithms, including generalized rank annihilation, bilinear least squares combined with residual bilinearization and multivariate curve resolution-alternating leastsquares. A set of experimental data was also studied, in which calibration was performed with fluorescence excitation-emission matrices for samples containing mixtures of chrysene (the analyte of interest) and benzopyrene (which produced strong inner filter effect across the useful wavelength range). Prediction was made on validation samples with a qualitative composition similar to the calibration set, and also on test samples containing an unexpected component (pyrene). In this latter case, U-PLS/RBL showed a unique success for the analysis of the calibrated component chrysene, achieving the useful second-order advantage.