Date of Award

2025

Document Type

Open Access Master's Thesis

Degree Name

Master of Science in Chemistry (MS)

Administrative Home Department

Department of Chemistry

Advisor 1

Rudy L. Luck

Committee Member 1

Marina Tanasova

Committee Member 2

Loredana Valenzano-Slough

Abstract

Within the field of biochemistry, there has been a significant development of ratiometric fluorescent probes designed for mitochondrial pH detection; however, many of these probes have emissions within the visible spectrum and require long synthesis times with low-yield reactions. However, in recent years, microwave-assisted syntheses of organic molecules have led to significant increases in reaction yields and quicker reaction times. In this work, two common reactions in probe synthesis, Heck coupling, and Knoevenagel condensations are investigated using microwave-assisted synthesis. It was found the low-concentration Heck coupling reactions using palladium (II) acetate led to extremely efficient Heck coupling. Also, a novel method for completing Knoevenagel condensations with multiple reactive sites with high selectively and increased reaction yields using microwave-assisted synthesis was completed. Moreover, two novel ratiometric pH-sensing probes, AH+ and BH+, were developed; both probes have emissions within the near-infrared range, with AH+ featuring ratiometric emission bands at 725 nm and 820 nm. Probe AH+ was evaluated for mitochondrial pH changes in living HeLa cells via fluorescence emission measurements of both ratiometric peaks. This probe was also used to measure the pH response within mitochondria with respect to different stimuli, such as starvation, oxidative stress, and hypoxia. Probe AH+ was also used to evaluate the pH effects of Metformin, a commercially available drug used in the treatments of diabetes, on mitochondrial pH, which allowed for the visualization of Metformin’s action within the intermembrane space of mitochondria.

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