Excitation-emission matrix data

Room-temperature fluorescence (RTF) has been used to determine the emission characteristics of a wide variety of materials relative to the wavelengths of selected Fraunhofer lines in support of the Fraunhofer luminescence detector remote-sensing instrument. RTF techniques are now used in the compilation of excitation-emission-matrix (EEM) fluorescence "signatures" of materials. The spectral data are collected with a Perkin-Elraer MPF-44B Fluorescence Spectrometer interfaced to an Apple 11+ personal computer. EEM fluorescence data can be displayed as 3-D perspective plots, contour plots, or "color-contour" images. The integrated intensity for selected Fraunhofer lines can also be directly extracted from the EEM data rather than being collected with a separate procedure. Fluorescence, chemical, and mineralogical data will be statistically analyzed to determine the probable physical and/or chemical causes of the fluorescence. 

There are several other chemometric approaches to calibration transfer that will only be mentioned in passing here. An approach based on finite impulse response (FIR) filters, which does not require the analysis of standardization samples on any of the analyzers, has been shown to provide good results in several different applications.81 Furthermore, the effectiveness of three-way chemometric modeling methods for calibration transfer has been recently discussed.82 Three-way methods refer to those methods that apply to A -data that must be expressed as a third-order data array, rather than a matrix. Such data include excitation/emission fluorescence data (where the three orders are excitation wavelength, emission wavelength, and fluorescence intensity) and GC/MS data (where the three orders are retention time, mass/charge ratio, and mass spectrum intensity). It is important to note, however, that a series of spectral data that are continuously obtained on a process can be constructed as a third-order array, where the three orders are wavelength, intensity, and time. 

There are two competing and equivalent nomenclature systems encountered in the chemical literature. The description of data in terms of ways is derived from the statistical literature. Here a way is constituted by each independent, nontrivial factor that is manipulated with the data collection system. To continue with the example of excitation-emission matrix fluorescence spectra, the three-way data is constructed by manipulating the excitation-way, emission-way, and the sample-way for multiple samples. Implicit in this definition is a fully blocked experimental design where the collected data forms a cube with no missing values. Equivalently, hyphenated data is often referred to in terms of orders as derived from the mathematical literature. In tensor notation, a scalar is a zeroth-order tensor, a vector is first order, a matrix is second order, a cube is third order, etc. Hence, the collection of excitation-emission data discussed previously would form a third-order tensor. However, it should be mentioned that the way-based and order-based nomenclature are not directly interchangeable. By convention, order notation is based on the structure of the data collected from each sample. Analysis of collected excitation-emission fluorescence, forming a second-order tensor of data per sample, is referred to as second-order analysis, as compared with the three-way analysis just described. In this chapter, the way-based notation will be arbitrarily adopted to be consistent with previous work. 

Molecular fluorescence spectroscopy is a commonly employed analytical method that is sensitive to certain chemical properties of FA (9-13). Fulvic acid s molecular fluorescence is principally due to conjugated unsaturated segments and aromatic moieties present in the macromolecule (14). Several types of fluorescence spectra can be measured, including an excitation emission matrix or total luminescence spectrum, constant offset synchronous fluorescence, excitation spectra, and emission spectra, furnishing the researcher with useful data. The ability to resolve and select multiple fluorescent species makes these approaches extremely useful for studying FA relative to its chemical reactivity. 

We developed two kinds of multidimensional fluorescence spectroscopic systems the time-gated excitation-emission matrix spectroscopic system and the time- and spectrally resolved fluorescence microscopic system. The former acquires the fluorescence intensities as a function of excitation wavelength (Ex), emission wavelength (Em), and delay time (x) after impulsive photoexcitation, while the latter acquires the fluorescence intensities as a function of Em, x, and spatial localization (%-, y-positions). In both methods, efficient acquisition of a whole data set is achieved based on line illumination by the laser beam and detection of the fluorescence image by a 2D image sensor, that is, a charge-coupled device (CCD) camera. 

Figure 32.1 (a) A schematic illustration of time-gated excitation-emission matrix spectroscopy, (b) A typical example of the 3D fluorescence data measured for Rhodamine 590 in ethanol... 

Summarizing this section, we developed the time-gated excitation-emission matrix spectroscopic system and applied it to the decomposition of a mixed solution of a number of fluorescent dyes. We demonstrated that our approach, which was based on unique optical configuration, efficient acquisition of a multidimensional data set, and decomposition of unknown fluorescent components by using the PARAFAC model, was effective for the analysis ofunknown multi-component targets. 

Another type of luminescence spectrum is shown in Figure I5 h. The total luminescence spectrum is cither a three-dimensional representation or a contour plot. Both simultaneously show the luminescence signal as a function of excitation and emission wavelengths. Such data aie often called an excitation-emission matrix. Although the total luniinescence spectrum can be obtained on u normal coni])U(cri/ed instrument, it can be acquired more rapidly with array-detector-based systems (see next seel ion). 

It is often convenient to format multidimensional fluorescence data in the form of a matrix of intensities called the excitation-emission matrix (EEM). The EEM can be represented as the vector product of the excitation and emission spectra. Each EEM has the following characteristics ... 

Total luminescence spectroscopy (TLS) is the simultaneous measurement of excitation, emission and intensity wavelengths of compound fluorophores [140-142]. This technique is mainly used for large ceU numbers in aqueous suspensions. In TLS the distinct fluorescence data that is generated from a three-dimensional matrix or excitation-emission matrix (EEM) of a specific microorganism is used for identification. Compared to two-dimensional emission spectra, this technique is highly sensitive and selective [143]. 

Although colorimetric methods were the earliest to be used for pesticide analysis [203], competitive spectroscopic methodologies for the determination of these pollutants were not developed until the last decade. The spectroscopic determination of several pesticides in mixtures has been the major hindrance, especially when their analytical characteristics are similar and their signals overlap as a result. Multivariate calibration has proved effective with a view to developing models for qualitative and quantitative prediction from spectroscopic data. Thus, partial least squares (PLS) and principal component regression (PCR) have been used as calibration models for the spectrofluorimetric determination of three pesticides (carbendazim, fuberidazole, and thiabendazole) [204]. A three-dimensional excitation-emission matrix fluorescence method has also been used for this purpose (Table 18.3) [205]. 

Xia, A. L., H. L. Wu, D. M. Fang, Y. J. Ding, L. Q. Hu, and R. Q. Yu. 2005. Alternating penalty trilinear decomposition algorithm for second-order calibration with application to interference-free analysis of excitation-emission matrix fluorescence data./. Chemom. 19 65-76. 

Any analytical data obtained by hyphenated instruments or by two-way spectroscopic techniques such as excitation-emission fluorescence spectroscopy are bilinear ones. The bilinear data matrix has a very useful property, namely the rank of such matrix obtained with any chemical mixture is equal to the number of chemical components in the mixture. Thus, theoretically, the rank of a data matrix of any pure chemical component is unit. It can be expressed by the product of two vectors ... 

Up to this point, regression has been restricted to two blocks of two-way data Y and X. In chemical analysis, however, a growing number of problems can be cast as three-way regression analysis. Consider the calibration of chemical constituents on the basis of their fluorescence excitation/emission spectrum or of gas chromatography/mass spectrometry (GC/MS) data. For each sample, two-dimensional measurements are available that constitute a three-way data array, X. This data array has to be related to sample concentrations of one, vector y, or several analytes, matrix Y. Cases can be imagined where even the matrix Y constitutes a three-way data array. 

The simultaneous fluorometric determination of PG and BHA in cosmetics has also been described using excitation-emission fluorescence matrix data, which were processed by applying the second-order calibration method based on self-weighted alternating normalized residue fitting algorithm [80]. The detection limit obtained for PG was 2.2 ng/mL. 

Subsequently, we reported the same experimental data as Merrill and Roberts but we have attributed the fluorescence to a (ir,ir ) transition rather than a (n,ir ) transition. We also pointed out that.the (n, ir ) state of PET is probably at higher energy than the 1 (tt.tt ) state Figure 2). We attributed the red shift in the fluorescence excitation and emission to aggregates of monomeric units fixed in specific geometry in the polymer matrix. 

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