Sarcosine Detection by Dichlorofluorescein as a Hydrogen Peroxide Probe
Department
Chemistry
Major
Chemistry
Research Advisor
Ma, Yinfa
Advisor's Department
Chemistry
Funding Source
Missouri S&T Opportunities for Undergraduate Research Experiences (OURE) Program; Department of Chemistry, Environmental Research Center
Abstract
Metabolites impart a significant importance to the understanding of biological reactions, and consequently, to the research of diagnostic and therapeutic techniques. Furthermore, there has been recent interest in metabolic levels present in urine. The detection of specific metabolites, however, presents certain analytical difficulties such as low or ambiguous specificity of the probe. This study proposes a new technique, utilizing oxidative, enzymatic production of hydrogen peroxide from the metabolite accompanied by dichlorodihydrofluorescein (DCFH). This probe displays high selectivity towards hydrogen peroxide, and coupled with high enzymatic specificity, forms an accurate method to measure metabolite levels. Sarcosine was used as a paradigm, by treatment of sarcosine oxidase to generate hydrogen peroxide, which was exposed to DCFH. Changes in fluorescence between initial and enzyme-exposed hydrogen peroxide levels were used to construct a standard curve from which actual urine samples were measured using this technique.
Biography
Casey was raised in the Lake of the Ozarks, MO where he attended the School of the Osage. In that time, he participated in his high school research class, putting out several papers, most notably that concerning the development of a computerized training method to demonstrate that the skill of absolute pitch could indeed be taught, earning him a chance to speak at many professional symposiums. He has taken these experiences with him to college, where he is currently participating in undergraduate research in Chemistry, and plans to make a career of it.
Research Category
Sciences
Presentation Type
Poster Presentation
Document Type
Poster
Award
Sciences poster session, First place
Location
Upper Atrium/Hallway
Presentation Date
06 Apr 2011, 9:00 am - 11:45 am
Sarcosine Detection by Dichlorofluorescein as a Hydrogen Peroxide Probe
Upper Atrium/Hallway
Metabolites impart a significant importance to the understanding of biological reactions, and consequently, to the research of diagnostic and therapeutic techniques. Furthermore, there has been recent interest in metabolic levels present in urine. The detection of specific metabolites, however, presents certain analytical difficulties such as low or ambiguous specificity of the probe. This study proposes a new technique, utilizing oxidative, enzymatic production of hydrogen peroxide from the metabolite accompanied by dichlorodihydrofluorescein (DCFH). This probe displays high selectivity towards hydrogen peroxide, and coupled with high enzymatic specificity, forms an accurate method to measure metabolite levels. Sarcosine was used as a paradigm, by treatment of sarcosine oxidase to generate hydrogen peroxide, which was exposed to DCFH. Changes in fluorescence between initial and enzyme-exposed hydrogen peroxide levels were used to construct a standard curve from which actual urine samples were measured using this technique.
