A flow-injection system is described which employs immobilized cholesterol esterase and cholesterol oxidase for the determination of total serum cholesterol. Hydrogen peroxide generated by the enzymatic reaction is measured directly at a home-made amperometric thin-layer flow-cell held at +600 mV vs Ag/AgCl. Serum samples are passed through an anion exchange resin to remove possible interferences prior to detection. The accuracy and precision of the system were evaluated with the use of certified reference materials for serum cholesterol and were found to conform to the recommended guidelines of the Laboratory Standardization Panel at the National Institutes of Health. / The use of sulfated polysaccharides as an affinity medium for the separation of the major classes of lipoproteins with subsequent determination of their cholesterol content was investigated. Several immobilization procedures for the covalent attachment of heparin to controlled-pore glass (CPG) were attempted. Separation of $ alpha$- (high-density) and $ beta$- (low- and very-low-density) lipoproteins was achieved on one of the heparin-CPG preparations and incorporation of this material into a chromatographic system with on-line determination of cholesterol was investigated. / Separation of $ alpha$- and $ beta$-lipoproteins was also achieved using sulfated dextran beads. A simple two-step procedure separates these two fractions in 10 minutes, and the cholesterol content is determined using the flow-injection system described above.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.74624 |
| Date | January 1991 |
| Creators | Carpenter, Alexis Anne |
| 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 | Doctor of Philosophy (Department of Chemistry.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001235332, proquestno: AAINN67668, Theses scanned by UMI/ProQuest. |
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