This thesis presents a quantitative protocol of proton nuclear magnetic resonance (1H NMR) that allows the determination of human amniotic fluid metabolite concentrations, which are then used in a metabonomic study to establish patient health during gestation. 1H NMR free inductive decays (FIDs) of 258 human amniotic fluid samples from a 500MHz spectrometer are acquired. Quantitative analyses methods in both the frequency- and time-domain are carried out and compared. Frequency-domain analysis is accomplished by integration of the metabolite peaks before and after the inclusion of a known standard addition of alanine. Time-domain analysis is accomplished by the direct exponential curve resolution algorithm (DECRA). Both techniques are assessed by applications to calibration biological solutions and a simulated data set. The DECRA method proves to be a more accurate and precise route for quantitative analysis, and is included in the developed protocol. Well-defined peaks of various components are visible in the frequency-domain 1H NMR spectra, including lactate, alanine, acetate, citrate, choline, glycine, and glucose. All are quantified with the proposed protocol. Statistical t-test and notched box and whisker plots are used to compare means of metabolite concentrations for diabetic and normal patients. Glucose, glycine, and choline are all found to correlate with gestational diabetes mellitus early in gestation. With further development, time-domain quantitative 1H NMR has potential to become a robust diagnostic tool for gestational health.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.100769 |
| Date | January 2007 |
| Creators | Botros, Lucy L. |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Master of Science (Department of Chemistry.) |
| Rights | © Lucy L. Botros, 2007 |
| Relation | alephsysno: 002590737, proquestno: AAIMR32665, Theses scanned by UMI/ProQuest. |
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