|dc.description.abstract||The study of carbohydrate structure and -metabolism has indicated the importance of
these compounds in the human body. Metabolic defects in the carbohydrate
metabolism, for example galactosemia and glycogen storage disease, result in serious
clinical complications as well as a direct influence on the energy production in the
body. The involvement of carbohydrates in multifactorial diseases has been
confirmed (Burke et al, 1996:609). The increase of all these diseases contributed to
the development of analytical techniques like mass spectrometry.
At present, the screening procedure for carbohydrate defects, is done by thin layer
chromatography. The method is fast and reasonable qualitative, but problems may
occur with definite identification as well as the influence of medication on the
analysis (Sewell, 1991:219). Mass spectrometry will be used complementary to thin
layer chromatography in this study. The MS-analysis are based on pre-column
derivatisation as well as MS-MS and LC-MS-MS detection and separation. Mono-,
di-, and oligosaccharides are derivatised by PMP (1-phenyl-3-methyl-2,5-
pyrazalone) and subjected to MS-MS and LC-MS-MS analysis (Rozakalis et 01,
2002). A complimentary analysis of ketose sugars is used in this study.
Derivatization of fructose and fructose containing metabolites are unfavourable.
Thus, an additional spectrophotometric method is implemented for the identification
of fructose and related compounds (colour reaction with a thryptamine reagent)
(Taylor, lWS:2 15).
This three way approach is an effective screening procedure for the definite
identification of carbohydrate metabolic diseases. It is indicated in this study that the
use of LC-MS-MS analysis may be confirmative to the TLC-results (especially in the
disorders like galactosemia). A variety of oligosaccharidoses, for example glycogen storage disease, a-mannosidosis, P-mannosidosis and fucosidosis, are successfully
identified by tandem mass spectrometry. An additional spectrophotometric analysis
confirms a possible fructosuria or fructose related defect. Through this approach,
effective identification of carbohydrates, which may be linked to a carbohydrate
metabolic disease or other carbohydrate related diseases, is made.||