A comparison of metabolite extraction and data manipulation methods for ¹H NMR spectroscopy of skeletal muscle using chicken breast tissue
Date of Award
Master of Science (MS)
Metabolism, Metabolism--Research, Proton magnetic resonance spectroscopy.
1H NMR has become an incredibly useful tool in the field of metabolomics. Many tissue types may be examined via 1H NMR in order to obtain a quantitative metabolic profile for an infinite number of phenotypes. In this study, I have tested three methods of metabolite extraction in conjunction with three methods of data analysis, in an attempt to determine which combination of the six methods (if any) are appropriate for extracting aqueous metabolites from skeletal muscle tissue for metabolite description and quantification via 1H NMR. Metabolite extraction methods tested the use of methanol/ chloroform/ water, methanol/ water, and HCl/ methanol as extraction reagents. Data analysis methods tested the use of adjustment to an internal standard (thymine), normalization to total protein content of the sample, and the combination of both. When comparing extraction methods alone, the methanol/ water method appears superior due to its simplicity, high concentration of extracted metabolites, and low relative standard errors. Combinations of extraction methods with data analysis methods showed that the methanol/ water extraction method used in conjunction with both thymine and total protein normalization was most optimal due to use of an internal standard, and low relative standard errors.
Underhill, Hannah, "A comparison of metabolite extraction and data manipulation methods for ¹H NMR spectroscopy of skeletal muscle using chicken breast tissue" (2023). Theses. 466.