Optical parameters measured via absorption spectroscopy and high-resolution fluorescence spectroscopy were used to characterize dissolved organic matter (DOM) in the springshed of Kings Bay, a spring-fed estuary located on Florida's Springs Coast. Over the past 40 years, springs supplying groundwater to Kings Bay have shown an increase in nitrate concentration. The overall goal of this project was to fingerprint wells and spring sites with elevated nitrogen concentrations using CDOM optical properties and establish relationships between nutrient and optical parameters. Samples were obtained from various sites: springs, Kings Bay surface (KBS), wells, coastal waters in and at the mouth of Crystal River (Coast) and lakes and rivers (LNR), during dry and wet seasons. The relationships between the environmental parameters and traditional optical parameters which provide insight into source characteristics were analyzed. Excitation emission matrix spectroscopy (EEMS) provided information about the concentration and chemical nature of organic matter in the study area. CDOM optical properties combined with salinity clearly separated the sources of fixed nitrogen in the Bay. Northern springs with elevated dissolved inorganic nitrogen (DIN) concentration had lower salinities and showed a presence of protein peaks. CDOM concentration was negatively correlated with total nitrogen (TN) and DIN, which suggests that these are subjected to anthropogenic influences. Humic peaks dominated the composition of the southern springs. CDOM concentrations were much higher than in the northern springs and there was a positive correlation between CDOM and both TN and DIN. These findings suggest that the fixed nitrogen in the southern springs is naturally occurring organic matter and the low concentrations may partially be a result of subsurface mixing of saltwater and freshwater in the aquifer. Thus, hypothesis testing showed that there was a significant difference between northern and southern springs Hypothesis testing also showed that there is a significant and unexpected positive relationship between CDOM and salinity studying Kings Bay, which is due to the low CDOM concentration in the springs discharging fresh water. This unique dataset also determined that the intercept of the mixing line was significantly different form zero. This indicates that CDOM is present and detectable at very low concentrations. Parallel Factor Analysis (PARAFAC) was used to evaluate CDOM composition from excitation emission matrix spectra (EEMs) and five components were identified: two humic, two marine humic, and one protein-like. The marine-like components, peak M, were produced in the marine environment and in meteoric groundwater. The study found a unique groundwater marker for coastal regions. Northern Kings Bay sites were characterized by a protein-like component, which has been associated with wastewater. Additional optical and environmental parameters were used in discriminate analysis, which successfully identified the CDOM markers for both natural and anthropogenic sources of nutrients in the environment. It is vital to improve the analysis of water, nutrients, and carbon from groundwater discharge into the coastal zone. Elevated DIN concentrations in groundwater are a widespread problem in Florida and over the past 30 years many spring waters have shown an increase in DIN concentrations. Nutrient discharge into delicate coastal areas can lead to ecological concerns. Investigating CDOM and nutrient distribution together can be a beneficial tool that can help differentiate sources from riverine/lacustrine, estuarine, marine, groundwater, and sewage impacted categories.

Philosophiae Doctor - PhD / Electroanalytical and optical properties of nanoscale materials are very important for biosensing applications as well as for understanding the unique one-dimensional carrier transport mechanism. One-dimensional semiconductor nanomaterials such as semiconductor quantum dots are extremely attractive for designing high-density protein arrays. Because of their high surfaceto-volume ratio, electro-catalytic activity as well as good biocompatibility and novel electron transport properties make them highly attractive materials for ultra-sensitive detection of biological macromolecules via bio-electronic or bio-optic devices. A genosensor or gene based biosensor is an analytical device that employs immobilized deoxyribonucleic acid (DNA) probes as the recognition element and measures specific binding processes such as the formation of deoxyribonucleic acid-deoxyribonucleic acid (DNA-DNA), deoxyribonucleic acid- ribonucleic acid (DNA-RNA) hybrids, or the interactions between proteins or ligand molecules with DNA at the sensor surface.In this thesis, I present four binary and two ternary-electrochemically and optically modulated selenide and telluride quantum dots, all synthesised at room temperature in aqueous media. Cationic gallium (Ga3+) synthesized in form of hydrated gallium perchlorate salt[Ga(ClO4)3.6H2O] from the reaction of hot perchloric acid and gallium metal was used to tailor the optical and electrochemical properties of the selenide and telluride quantum dots. The synthesized cationic gallium also allowed successful synthesis of novel water soluble and biocompatible capped gallium selenide nanocrystals and gallium telluride quantum dots. Cyclic voltammetric studies inferred that presence of gallium in a ZnSe-3MPA quantum dot lattice improved its conductivity and significantly increased the electron transfer rate in ZnTe-3MPA.Utraviolet-visible (UV-vis) studies showed that incorporation of gallium into a ZnSe-3MPA lattice resulted in a blue shift in the absorption edge of ZnSe-3MPA from 350 nm to 325 nm accompanied by decrease in particle size. An amphiphilic bifunctional molecule, 3-Mercaptopropionic acid (3-MPA) was used as a capping agent for all quantum dots. It was found that 3-MPA fully solubilised the quantum dots, made them stable, biocompatible, non agglomerated and improved their electron transfer kinetics when immobilized on gold electrodes.Retention of the capping agent on the quantum dot surface was confirmed by Fourier transform infrared spectroscopy (FTIR) which gave scissor type bending vibrations of C-H groups in the region 1365 cm-1 to 1475 cm-1, stretching vibrations of C=O at 1640 cm-1, symmetric and asymmetric vibrations of the C-H in the region 2850 cm-1 to 3000 cm-1 as well as stretching vibrations of –O-H group at 3435 cm-1. The particle size and level of non-agglomeration of the quantum dots was studied by high resolution transmission electron microscopy (HRTEM). The optical properties of the quantum dots were studied using UV-vis and fluorescence spectroscopic techniques.Quantum dot/nanocrystal modified gold electrodes were prepared by immersing thoroughly cleaned electrodes in the quantum dot/nanocrystal solution, in dark conditions for specific periods of time. The electrochemical properties of the modified electrodes were characterized by cyclic voltammetry (CV), square wave voltammetry (SWV), electrochemical impedance and spectroscopy (EIS). Six sensing platforms were then prepared using quantum dot/nanocrystal, one of which was used for detection of dopamine while the rest were used for detection of a DNA sequence related to 5-enolpyruvylshikimate-3-phosphate synthase, a common vector gene in glyphosate resistant transgenic plants.The first sensing platform, consisting of ZnSe-3MPA modified gold electrode (Au|ZnSe-3MPA) gave rise to a novel method of detecting dopamine in presence of excess uric acid and ascorbic acid. Using a potential window of 0 to 400 mV, the ZnSe-3MPA masked the potential for oxidation of uric and ascorbic acids, allowing detection of dopamine with a detection limit of 2.43 x 10-10 M (for SWV) and 5.65 x 10-10 M (for steady state amperometry), all in presence of excess uric acid (>6500 higher) and ascorbic acid (>16,000 times higher). The detection limit obtained in this sensor was much lower than the concentration of dopamine in human blood(1.31 x 10-9 M), a property that makes this sensor a potential device for detection of levels of dopamine in human blood.The other sensing platforms were prepared by bioconjugation of amine-terminated 20 base oligonucleotide probe DNA (NH2-5′-CCC ACC GGT CCT TCA TGT TC-3′) onto quantum dot modified electrodes with the aid of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). The prepared DNA electrodes were electrostatically hybridized with different sequences which included 5′-GAA CAT GAA GGA CCG GTG GG-3′ (complementary target), 5′-CATAGTTGCAGCTGCCACTG-3′ (non complementary target) and 5′-GATCATGAAGCACCGGAGGG-3′ (3-base mismatched target).The hybridization events were monitored using differential pulse voltammetry (DPV) and SWV by monitoring the guanine oxidation signal or using EIS by monitoring changes in the charge transfer resistance. The quantum dot genosensors were characterized by low detection limits (in the nanomolar range), long linear range (40 - 150 nM) and were able to discriminate among complementary, non-complementary and 3-base mismatched target sequences.