Harwood-Stamper, Annie Joann 
ORCID: 0000-0003-0077-3794
(2022).
Developing innovative analytical and computational methods for the study of human metabolism.
University of Birmingham.
Ph.D.
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Harwood-Stamper2022PhD.pdf
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Abstract
Currently, within untargeted metabolomics there are several limitations that are delaying the speed at which new knowledge can be gained. One of these is the analysis methods applied where only low sample volume/masses are available. Another is metabolite annotation and identification because two orthogonal data matches with matching data collected for chemical standards in-house are required for identification. Metabolomics is commonly applied within heath and exercise science and is extremely important to investigate the effects of exercise on the human body for health research, sports performance, and nutritional needs. There have been multiple studies conducted to study the differences in medium, high, and maximal intensity exercise, but none that have compared all three at once. Another area of health and exercise metabolomics that is currently limited is heat acclimation. As this is commonly applied to both military and athletes, allowing for optimal performance in hot climates, more understanding on how heat acclimation affects the human metabolome is required.
This thesis aims to develop and optimize UHPLC-MS methods to allow for a wider range of sample types to be analysed along with expanding the bottleneck area of metabolite annotation further. Aqueous reversed phase, lipidomic C18 reversed phase and HILIC assays were developed applying 1mm internal diameter (ID) columns and compared to commonly applied 2.1mm ID columns for the same assays. Following this, the 1mm ID assays were validated using porcine plasma, urine, and tear fluid samples. Results demonstrated that current 2.1mm ID assays are suitable for biofluid sample volumes typically collected in untargeted metabolomic studies. When applying more dilute samples, a 1mm ID column
becomes more advantageous. Results for the porcine tear fluid demonstrated that tear fluid is a metabolite rich sample type that is applicable for future metabolomics research.
Next, retention time mapping methods were developed and cross validated to allow for different RT libraries around the world to be able to be transferred between research groups. Results concluded that RT mapping methods proved that it is possible to transfer retention times between research groups, even when alternative UHPLC-MS instrumentation, column stationary phases and mobile phases were applied. An in-house lipids retention time library was constructed from regression models for each class with the aim to predict the retention time for lipids of the same class where chemical standards are not available.
This thesis also aims to expand the knowledge gap areas on how the human body reacts to heat acclimation and different intensities of exercise. Two different biological studies were conducted to investigate how the human metabolome responds to different intensities of exercise and heat
acclimation, respectively. Results showed that as the intensity of exercise increases, the body response changes as there is a larger requirement for energy and that heat acclimation influences biofluid metabolomes, with multiple pathways being perturbed across both days of condensed heat acclimation sessions.
| Type of Work: | Thesis (Doctorates > Ph.D.) | |||||||||||||||
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| Award Type: | Doctorates > Ph.D. | |||||||||||||||
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| Licence: | All rights reserved | |||||||||||||||
| College/Faculty: | Colleges (2008 onwards) > College of Life & Environmental Sciences | |||||||||||||||
| School or Department: | School of Biosciences | |||||||||||||||
| Funders: | Engineering and Physical Sciences Research Council | |||||||||||||||
| Subjects: | Q Science > QD Chemistry Q Science > QH Natural history > QH301 Biology |
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| URI: | http://etheses.bham.ac.uk/id/eprint/12500 |
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