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High pressure and microwave assisted generation and pyrolysis-GCMS analysis of glycated proteins

The extent of denaturation and glycation of lysozyme and BSA with the application of high hydrostatic pressure (HHP) at 400 MPa at 30°C from 8 to 48 hours and focused microwave irradiation at 50°C under varying microwave power and from 10 to 60 minutes was investigated in the presence and absence of D-glucose. The HHP treatment caused 10 to 20% denaturation of lysozyme whereas microwave irradiation caused 20 to 40% denaturation, with more destruction to the lysozyme in the presence of glucose compared to the control. The extent of glycation was also higher with the high pressure samples, causing 60% glycation upon 8 hours of high pressure exposure, but decreasing to around 40% thereafter. Microwave irradiation brought about 40% glycation to the lysozyme samples upon 20 min of irradiation. BSA, on the other hand, was more susceptible to damage by high energy exposures. BSA samples were denatured to a greater extent compared to lysozyme, up to 80% upon the prolonged exposures, but in all treatments, glucose seemed to act as a protectant contrary to the case of lysozyme. The extent of glycation detected was also minimal, ranging from 8 to 20%. / The feasibility of analyzing glycated proteins using pyrolysis-GC/MS was also investigated. Taking advantage of the formation of a diagnostic marker---2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one---upon pyrolysis of glycated proteins, the intensity of this peak was used to correlate the extent of glycation. The intensity of this peak in the pyrograms of glycated lysozymes was found to increase linearly with increasing incubation times and subsequently with the sugar loads of the glycated lysozyme. In addition, using the pyrograms as unique fingerprints, the extent of structural changes between modified and unmodified proteins were also assessed.

Identiferoai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.79033
Date January 2002
CreatorsLi, Pik Kei, 1978-
ContributorsYaylayan, V. A. (advisor)
PublisherMcGill University
Source SetsLibrary and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada
LanguageEnglish
Detected LanguageEnglish
TypeElectronic Thesis or Dissertation
Formatapplication/pdf
CoverageMaster of Science (Department of Food Science and Agricultural Chemistry.)
RightsAll items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated.
Relationalephsysno: 001985735, proquestno: AAIMQ88248, Theses scanned by UMI/ProQuest.

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