Chronic, noncommunicable diseases such as chronic fatigue syndrome (also known as
myalgic encephalomyelitis) are rapidly becoming a worldwide epidemic that profoundly
affects public health and productivity. Chronic fatigue syndrome (CFS) is characterised by
severe and debilitating fatigue and although its etiology is still unknown, recent studies have
found considerable evidence that mitochondrial dysfunction and oxidative stress might be
responsible for the underlying energy deficit in these patients. Adenine and pyridine
nucleotides could be used as potential biomarkers for energy related disorders such as
chronic fatigue syndrome because of their various functions in the energy and redox
pathways.
The first part of this study focussed on developing a liquid chromatography electrosprayionisation
tandem mass spectrometry (LC-ESI-MS/MS) method for the quantification of
these nucleotides in blood samples. Due to the instability of nucleotides in biological
matrices it was also necessary to find a suitable extraction method that would be able to stop
enzymatic activity via protein precipitation. Out of the four extraction methods investigated
during this study, deproteinisation of whole blood samples with perchloric acid produced the
highest nucleotide abundances. Although nucleotide standards were found to be stable in
perchloric acid, nucleotide levels in blood samples were not stabilised by addition of
perchloric acid.
The second part of this study consisted of measuring the nucleotide levels in blood samples
of controls and possible CFS patients in order to test the proof of concept of the new LCESI-
MS/MS method. Despite changes in the nucleotide levels due to perchloric acid and
problems with nucleotide instability, it was still possible to distinguish between the two
groups based on the results obtained with the new LC-ESI-MS/MS method.
The newly developed LC-ESI-MS/MS method proved to be reliable and adequate for
nucleotide quantification in whole blood samples, thus the aim of this study was achieved. / MSc (Biochemistry), North-West University, Potchefstroom Campus, 2014
| Identifer | oai:union.ndltd.org:NWUBOLOKA1/oai:dspace.nwu.ac.za:10394/10768 |
| Date | January 2014 |
| Creators | Moolman, Chantalle |
| Source Sets | North-West University |
| Language | English |
| Detected Language | English |
| Type | Thesis |
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