Novel Analytical Methods for Improved Analysis of Biological Compounds
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| Language: | English |
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The Ohio State University / OhioLINK
2015
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| Online Access: | http://rave.ohiolink.edu/etdc/view?acc_num=osu1447408723 |
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ndltd-OhioLink-oai-etd.ohiolink.edu-osu1447408723 |
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NDLTD |
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English |
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Chemistry Analytical Chemistry High-performance liquid chromatography Enhanced-fluidity liquid chromatography Surface-assisted laser desorption ionization Matrix-enhanced laser desorption ionization Mass spectrometry |
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Chemistry Analytical Chemistry High-performance liquid chromatography Enhanced-fluidity liquid chromatography Surface-assisted laser desorption ionization Matrix-enhanced laser desorption ionization Mass spectrometry Beres, Martin Joseph Novel Analytical Methods for Improved Analysis of Biological Compounds |
| author |
Beres, Martin Joseph |
| author_facet |
Beres, Martin Joseph |
| author_sort |
Beres, Martin Joseph |
| title |
Novel Analytical Methods for Improved Analysis of Biological Compounds |
| title_short |
Novel Analytical Methods for Improved Analysis of Biological Compounds |
| title_full |
Novel Analytical Methods for Improved Analysis of Biological Compounds |
| title_fullStr |
Novel Analytical Methods for Improved Analysis of Biological Compounds |
| title_full_unstemmed |
Novel Analytical Methods for Improved Analysis of Biological Compounds |
| title_sort |
novel analytical methods for improved analysis of biological compounds |
| publisher |
The Ohio State University / OhioLINK |
| publishDate |
2015 |
| url |
http://rave.ohiolink.edu/etdc/view?acc_num=osu1447408723 |
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AT beresmartinjoseph novelanalyticalmethodsforimprovedanalysisofbiologicalcompounds |
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1719439103561302016 |
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ndltd-OhioLink-oai-etd.ohiolink.edu-osu14474087232021-08-03T06:33:43Z Novel Analytical Methods for Improved Analysis of Biological Compounds Beres, Martin Joseph Chemistry Analytical Chemistry High-performance liquid chromatography Enhanced-fluidity liquid chromatography Surface-assisted laser desorption ionization Matrix-enhanced laser desorption ionization Mass spectrometry The work contained within this dissertation focuses on innovative technologies in the field of analytical chemistry, particularly within high-performance liquid chromatography (HPLC) and mass spectrometry (MS). Enhanced-fluidity liquids (EFL), which have low viscosity and high diffusivity, were studied as alternative mobile phases in mixed-mode hydrophilic interaction strong ion-exchange chromatography (HILIC/SCX). Additionally, these mobile phases were evaluated as environmentally friendly alternatives to traditional HILIC solvents in gradient separations. Finally, electrospun nanofibrous materials with high surface area to volume ratios were assessed as substrates in surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS).The potential of enhanced-fluidity liquid chromatography (EFLC) HILIC/SCX was explored, using amino acids as analytes. EFL mobile phases were prepared by adding liquefied CO<sub>2</sub> to methanol:water (MeOH:H<sub>2</sub>O) mixtures, which increases the diffusivity and decreases the viscosity of the mixture. The optimized chromatographic performance of these MeOH:H<sub>2</sub>O:CO<sub>2</sub> EFL mixtures was compared to traditional acetonitrile:water (ACN:H<sub>2</sub>O) and MeOH:H<sub>2</sub>O liquid chromatography (LC) mobile phases. MeOH:H<sub>2</sub>O:CO<sub>2</sub> mixtures offered higher efficiencies and resolution of the ten amino acids relative to the MeOH:H<sub>2</sub>O LC mobile phase, and decreased the required isocratic separation time by a factor of two relative to the ACN:H<sub>2</sub>O LC mobile phase. Large differences in selectivity were also observed between the EFLC and LC mobile phases. Retention mechanism studies revealed that the EFLC mobile phase separation was governed by a mixed-mode retention mechanism of HILIC/SCX. On the other hand, separations with ACN:H<sub>2</sub>O and MeOH:H<sub>2</sub>O LC mobile phases were strongly governed by only one retention mechanism, either HILIC or SCX, respectively. EFLC was then evaluated for “green” HILIC separations. The impact of CO<sub>2</sub> addition to a MeOH:H<sub>2</sub>O mobile phase was studied as an alternative to traditional ACN:H<sub>2</sub>O HILIC mobile phases, while also optimizing buffer type, ionic strength, and pH. Using EFLC mixtures, a separation of 16 RNA nucleosides/nucleotides was achieved in 16 minutes with greater than 1.3 resolution for all analyte pairs. By using a reverse CO<sub>2</sub> gradient, analysis time was reduced by over 100% in comparison to isocratic conditions. The optimal separation using MeOH:H<sub>2</sub>O:CO<sub>2</sub> mobile phases was also compared to that using MeOH:H<sub>2</sub>O and ACN:H<sub>2</sub>O mobile phases. Based on the chromatographic performance parameters (efficiency, resolution, and speed of analysis) and the overall environmental impact of the mobile phase mixtures, MeOH:H<sub>2</sub>O:CO<sub>2</sub> mixtures were preferred to ACN:H<sub>2</sub>O or MeOH:H<sub>2</sub>O mobile phases for the separation of mixtures of these RNA nucleosides and nucleotides. Finally, electrospun nanofibrous substrates were studied for the improvement of SALDI-MS analysis of large molecular weight proteins and polymers without the use of a chemical matrix. Various polymers (including polyacrylonitrile, polyvinyl alcohol, and SU-8 photoresist) and carbon substrates were examined. SALDI analysis using these substrates eliminated “sweet spot” formation typically seen in matrix-assisted laser desorption/ionization (MALDI), which lead to greater shot-to-shot reproducibility. The fiber diameter of these substrates played a significant role in the quality of the mass spectra generated, with smaller fiber diameter yielding higher signal to noise ratio (S/N). Additionally, the degree of pyrolysis also impacted the degree of fragmentation and overall S/N for the prepared carbon substrates. 2015 English text The Ohio State University / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=osu1447408723 http://rave.ohiolink.edu/etdc/view?acc_num=osu1447408723 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws. |