1 characterization of bsn 175 : a sugar substitute drug

BSN175 is a novel sugar substitute drug intended to treat Prader-Willi syndrome, a condition that affects thousands of children worldwide. Characterized by weakness in infancy and obesity, behavioral issues, and an insatiable appetite from the age of two onwards, this condition is all-consuming and can be devastating to those experiencing it and their families. Type II diabetes frequently accompanies this obesity. While there is no cure for Prader-Willi, there are ways to manage the symptoms. Sugar substitutes are used to reduce caloric intake and sate those who cannot control their appetites. BSN175 is one such sugar substitute drug that has been investigated for use in treatment of type II diabetes. To provide quantifiable, reproducible analysis of BSN175, the active pharmaceutical ingredient, D-tagatose, needs to be easily identified and differentiated from its decomposed form and other possible impurities. Infrared and proton NMR spectroscopy will be applied to the drug to build a package that can be used to determine the identity and purity of D-tagatose to meet FDA regulations for quality and safety. 2 THE DESIGN AND FABRICATION OF COMPONENTS FOR A STAGE-TOP HIGH-THROUGHPUT TIRF MICROSCOPY MODULE Cassandra Almasri, Senior, Chemistry, University of Kentucky Chris Richards, Chemistry, University of Kentucky Jason DeRouchey, Chemistry, University of Kentucky Abstract: Total Internal Reflection Fluorescence Microscopy (TIRFM) is a technique used to directly visualize the fluorophores on the surface of a given sample. TIRFM is best known for its applications used to visualize processes occurring in the plasma membranes of cells; however, it has a wide variety of applications beyond the visualization of biomolecules. TIRFM works by using fiber optics to project a laser beam of desired wavelength onto the surface of the sample at an incident angle. By shining light at an incident angle greater than the critical angle, light will be reflected from the sample and only the surface of the sample will be illuminated and analyzed. Total Internal Reflection Fluorescence Microscopy (TIRFM) is a technique used to directly visualize the fluorophores on the surface of a given sample. TIRFM is best known for its applications used to visualize processes occurring in the plasma membranes of cells; however, it has a wide variety of applications beyond the visualization of biomolecules. TIRFM works by using fiber optics to project a laser beam of desired wavelength onto the surface of the sample at an incident angle. By shining light at an incident angle greater than the critical angle, light will be reflected from the sample and only the surface of the sample will be illuminated and analyzed. Current methods for analyzing samples using TIRFM are directed towards single sample analysis. This becomes particularly challenging in fields that are interested in carrying out reliable studies with different samples and/or different timepoints. The purpose of this research is to propose and develop a system that allows for widespread analysis of samples using high-throughput TIRF. In this project we suggest HT-TIRF would employ a prism-based stage top design compatible with a 96 well plate. This design will allow for HTTIRF on a fluorescent microscope by shining a laser onto a prism located directly under the wells and allowing a propagated evanescent illumination to extend onto the wells that will be analyzed. The efficiency and reliability of the device and corresponding fiber optic system will be further investigated by imaging several test samples. 3 QUANTIFYING DIFFUSION OF A MODEL ANALYTE SET VIA CAPILLARY ELECTROPHORESIS Corbin Arrasmith, Junior, Chemistry, Northern Kentucky University Charlisa Daniels, Chemistry, Northern Kentucky University Abstract: The goal of this project was to further understand the relationships between the temperature, analyte, and concentration of the analyte within a capillary. This was investigated using Capillary Electrophoresis (CE) on the PAH series (Naphthalene through Chrysene). This series was selected based on their use in previous studies. In addition further analysis’s on the GMA:EDMA polymers was completed along with Scanning Electron Microscope (SEM) images to show the pores size of the polymer and how they change based of GMA:EDMA ratios and porogen size. The goal of this project was to further understand the relationships between the temperature, analyte, and concentration of the analyte within a capillary. This was investigated using Capillary Electrophoresis (CE) on the PAH series (Naphthalene through Chrysene). This series was selected based on their use in previous studies. In addition further analysis’s on the GMA:EDMA polymers was completed along with Scanning Electron Microscope (SEM) images to show the pores size of the polymer and how they change based of GMA:EDMA ratios and porogen size. 4 SEPARATION AND ANALYSIS OF HEPARAN SULFATE TETRASSACHARIDE MIXTURES USING CAPILLARY ELECTROPHORESIS, AND ORBITRAP MASS SPECTROME John Attelah, Junior, Chemistry, Berea College Patience Sanderson, Chemistry, University of Georgia Morgan Stickeney, Chemistry, University of Georgia Jonathan Amster, Chemistry, University of Georgia Abstract: Glycosaminoglycans (GAGs) are complex polysaccharides, expressed at the cell surface and in the extracellular matrix, which comprise the carbohydrates portion of proteoglycans and are found in a variety of organisms ranging from bacteria to humans. GAGs participate in a number of significant biological processes, such as cell-cell and cell-matrix signaling at the origin of a variety of physiological and pathological functions such as embryonic development, cell adhesion, the regulation of biochemical pathways, cell growth and differentiation, homeostasis, and the mediation of inflammatory reactions. They have also been implicated in the initial step of some pathogenic infections and have been observed to undergo some alteration in some tumor growth. There are different types of GAGs which include Chondroitin Sulfates(CS), Dermatan Sulfates (DS), Keratan Sulfates(KS), Heparin and Heparan Sulfates(HS). This research focused on separation and analysis of heparan sulfate tetrassacharide mixture using Capillary Electrophoresis and Orbitrap Mass Spectrometry. Glycosaminoglycans (GAGs) are complex polysaccharides, expressed at the cell surface and in the extracellular matrix, which comprise the carbohydrates portion of proteoglycans and are found in a variety of organisms ranging from bacteria to humans. GAGs participate in a number of significant biological processes, such as cell-cell and cell-matrix signaling at the origin of a variety of physiological and pathological functions such as embryonic development, cell adhesion, the regulation of biochemical pathways, cell growth and differentiation, homeostasis, and the mediation of inflammatory reactions. They have also been implicated in the initial step of some pathogenic infections and have been observed to undergo some alteration in some tumor growth. There are different types of GAGs which include Chondroitin Sulfates(CS), Dermatan Sulfates (DS), Keratan Sulfates(KS), Heparin and Heparan Sulfates(HS). This research focused on separation and analysis of heparan sulfate tetrassacharide mixture using Capillary Electrophoresis and Orbitrap Mass Spectrometry. 5 FLUOROAMIDE-DIRECTED FLUORINATION OF UNACTIVATED C–H BONDS Jenna Bingham, Senior, Chemistry, Indiana University Brian Groendyke, Chemistry, Indiana University Silas Cook, Chemistry, Indiana University Abstract: Organofluorine compounds possess unique and desirable properties with applications spanning the pharmaceutical, agricultural, and materials industries. Despite being ideal transformations, methods to selectively fluorinate Csp3-H bonds are still scarce and in need of development. Previous work in the Cook Group demonstrated that N-fluoro-2-methylbenzamides selectively transfer fluorine upon reaction with iron (II) triflate to provide the benzyl fluoride in yields up to 93%. Attempts to change from the rigid 2methylbenzamide class to linear alkyl systems caused the yields to plummet. This work presents our latest efforts in the optimization of the fluorine transfer reaction for open-chain, unactivated systems. Organofluorine compounds possess unique and desirable properties with applications spanning the pharmaceutical, agricultural, and materials industries. Despite being ideal transformations, methods to selectively fluorinate Csp3-H bonds are still scarce and in need of development. Previous work in the Cook Group demonstrated that N-fluoro-2-methylbenzamides selectively transfer fluorine upon reaction with iron (II) triflate to provide the benzyl fluoride in yields up to 93%. Attempts to change from the rigid 2methylbenzamide class to linear alkyl systems caused the yields to plummet. This work presents our latest efforts in the optimization of the fluorine transfer reaction for open-chain, unactivated systems. 6 SYNTHESIS OF MULTICOLOR FLUORESCENT LIGANDS FOR NICOTINIC ACETYLCHOLINE RECEPTORS Alexandra Davis, Senior, Chemistry, Indiana State University Richard Fitch, Chemistry, Indiana State University Abstract: Nicotinic acetylcholine receptors (nAChR) are ligand gated ion channels sensitive to the neurotransmitter acetylcholine and serve critical roles in preand postsynaptic neurotransmission in the central and peripheral nervous systems. Fluorescent ligands for nAChR can be used to locate nicotinic receptors in isolated tissues and cultured cells. Our design of fluorescent ligands include a ligand motif (A-84543), a hydrophilic linker, and terminal fluorophore. We are examining a variety of fluorophores for multilabeling applications in fluorescent microscopy, flow cytometry and binding/functional assays for nAChR. Our results to date will be described. Nicotinic acetylcholine receptors (nAChR) are ligand gated ion channels sensitive to the ne