Hadamard-Transform Fluorescence Excitation-Emission-Matrix Spectroscopy: Rapid 3-Dimensional Fluorescence Data for Analytical and Industrial Applications
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Authors
Ferguson, Travis
Date
Type
thesis
Language
eng
Keyword
Chemistry , Spectroscopy , Hadamard Transform , Fluorescence , Analytical Chemistry , Physical Chemistry , Hadamard , Multiplexing , Excitation Emission Matrix Spectroscopy , EEM Spectroscopy , HPLC
Alternative Title
Abstract
Fluorescence spectroscopy is a very sensitive and selective analytical technique that can be used to
monitor and quantify a sample. The method is non-destructive and creates no chemical waste. In
conventional fluorescence spectroscopy, a sample is excited with a single wavelength of light and the
emitted spectrum is collected. The conventional method is well suited for simple samples and sample
mixtures, but it fails when several fluorophores emit similar spectra or have a low absorption cross
section at the excitation wavelength. Excitation-emission-matrix (EEM) fluorescence spectroscopy
addresses both shortcomings by using many excitation wavelengths and recording a complete emission
spectrum at each excitation wavelength. This creates a two-dimensional spectrum, which may be
displayed as a contour plot or “heat map”. Fluorophores produce “hills” in these EEM spectra and may be
identified and quantified using multivariate data analysis methods.
Conventional EEM spectrometers scan the excitation light sequentially through a region of the UV-Vis
spectrum while recording the emission spectra of the fluorophore with a spectrometer; this process may
take up to an hour for a well-resolved spectrum. Not surprisingly, EEM spectroscopy has previously not
been used for kinetic studies or the detection of transient species and intermediates. In this thesis, a new
Hadamard-transform excitation-emission-matrix (HT-EEM) spectrometer is described and developed,
which reduces the data acquisition time from 30-60 minutes to under 140 milliseconds. A new data
acquisition algorithm (Super-Cycle algorithm) was applied to a HT-EEM dataset, and it demonstrated an
even higher time resolution at less than 20 ms/spectrum. As a result, analytical problems that were
intractable only a few years ago are now within reach. For example, with our rapid analysis time we can
perform on-line and real-time analysis of samples, such as turbine oils, and check their molecular
composition and quality. Our new HT-EEM technique was also applied to existing analytical methods
such as HPLC separation, where we can record the EEM spectrum of the different fluorophores within a
sample.
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Attribution-NonCommercial-ShareAlike 3.0 United States
ProQuest PhD and Master's Theses International Dissemination Agreement
Intellectual Property Guidelines at Queen's University
Copying and Preserving Your Thesis
This publication is made available by the authority of the copyright owner solely for the purpose of private study and research and may not be copied or reproduced except as permitted by the copyright laws without written authority from the copyright owner.
Attribution-NonCommercial-ShareAlike 3.0 United States