The analysis of biological fluids for acylcarnitines

Kelly, Barbara M. (2000). The analysis of biological fluids for acylcarnitines. PhD thesis The Open University.



This thesis outlines the development and evolution of a method for the analysis of biological fluids for acylcamitines. These compounds are of interest due to their role in the metabolism of fatty acids and their use as indicators of metabolic defects, which have been associated with SIDS (Sudden Infant Death Syndrome).

These compounds are zwitterionic and thermally labile in nature and would not traditionally lend themselves to gas chromatographic - mass spectrometric (GC-MS) analysis. Described in this thesis is a method whereby lactonisation of the acylcamitine facilitates the volatility required for GC analysis while retaining the side chain identifier to the original acylcamitine for mass spectrometric identification. Using this method it has been possible to characterize acylcamitines which are metabolic indicators, distinguish isomeric acylcamitines chromatographically and show diagnosis of a number of diseases from clinical samples.

Sample analysis has been demonstrated for both blood and urine. In the case of urine samples an example has been provided to illustrate the advances in analytical techniques available for these analyses, to show the stability of these compounds in the matrix and the wealth of diagnostic information which can be obtained for an individual sample. The development of a method for the analysis of acylcamitines from blood spots has been detailed. The application of this methodology to a number of disorders of fatty acid metabolism has been illustrated. The extension of the technique to the analysis of dicarboxylic acylcamitines has also been investigated.

Finally with advances in modem technology the area of eléctrospray mass spectrometry has been evaluated for the analysis of samples without lactonisation. Results from this investigation are presented.

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