Fluorescence modeling in a multicomponent system

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Abstract
An extensive fluorescence database for binary tyrosinetryptophan mixtures utilizing 280 nm excitation was collected. The database spanned three orders of magnitude (10−6M–10−3M) and covered all compositions within this range. A generalized model for describing the multicomponent fluorescence signals as a function of emission wavelength, excitation wavelength, and sample composition was derived. A geometric integral that contained all the geometric factors affecting fluorescence was introduced; thus the model was applicable to various configurations, including the three used in this study: an NADH probe, a backscatter laser‐induced fluorescence setup, and a commercial spectroflurometer. A correction factor was proposed that allowed linearization of the fluorescence signals with respect to fluorophore concentrations. The effect of the water Raman on fluorescence spectra was also modeled. The model contains only two wavelength‐dependent parameters for each of the components present in a sample, one specifying absorption of the excitation energy and the other specifying the species' fluorescence tendency. These wavelength‐dependent parameters were correlated with polynomials. The average prediction error at each wavelength was 10–20%, a major portion of which was attributed to experimental uncertainties.