Department of Chemistry, Mississippi State University, Mississippi State, Mississippi 39762, United States.
Department of Mathematical Sciences, DePaul University, Chicago, Illinois 60604, United States.
Anal Chem. 2022 May 17;94(19):7123-7131. doi: 10.1021/acs.analchem.2c01131. Epub 2022 May 4.
The popular textbook and literature model (λ,λ) = or its variants for correlating the sample absorption and fluorescence often fails even for the simplest samples where the fluorophore is the only light absorber. Reported is a first-principle model (λ,λ) = for correlating the sample fluorescence measured with a conventional spectrofluorometer and its UV-vis absorbance quantified with a conventional UV-vis spectrophotometer. This model can be simplified or expanded for a variety of fluorescence analyses. First, it enables curve-fitting fluorescence intensity as a function of the fluorophore or sample absorbance over a sample concentration range impossible with existing models. Second, it provides the theoretical foundation for an inner-filter-effect (IFE)-correction method developed earlier and explains mathematically the linearity between the IFE-corrected fluorescence and the fluorophore concentration or absorbance. Third, this model can be expanded for quantitative mechanistic studies of fluorescence intensity variations triggered by stimuli treatments. One demonstrated example is to quantify temperature effects on the emission-wavelength-specific and total fluorescence quantum yield of anthracene. We expect that this first-principle model will be broadly adopted for both student education that promotes evidence-based learning and a variety of fluorescence applications where disentangling sample absorption and emission are critical for reliable data analysis.
流行的教科书和文献模型（λ，λ）= 或其变体用于相关联的样本吸收和荧光，即使对于最简单的样本，其中荧光团是唯一的光吸收体，也经常失败。本文报道了一种用于关联传统荧光分光光度计测量的样品荧光和传统紫外-可见分光光度计量化的样品紫外-可见吸光度的第一性原理模型（λ，λ）= 。该模型可简化或扩展用于各种荧光分析。首先，它使我们能够在现有模型不可能的样品浓度范围内拟合荧光强度作为荧光团或样品吸光度的函数。其次，它为之前开发的内滤效应（IFE）校正方法提供了理论基础，并从数学上解释了 IFE 校正后的荧光与荧光团浓度或吸光度之间的线性关系。第三，该模型可扩展用于定量研究刺激处理引发的荧光强度变化的机制。一个演示示例是量化温度对蒽的发射波长特异性和总荧光量子产率的影响。我们期望该第一性原理模型将被广泛应用于促进基于证据学习的学生教育以及各种荧光应用，其中解耦样品吸收和发射对于可靠的数据分析至关重要。
