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Dried blood spot sampling method in determination of amino acids, keto acids, fatty acids and glutathione
Authors: Andrlová Lenka | Kanďár Roman
Year: 2016
Type of publication: ostatní - přednáška nebo poster
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
eng Dried blood spot sampling method in determination of amino acids, keto acids, fatty acids and glutathione In a comparison with classic liquid samples, dried blood spot (DBS) sampling method has several advantages. One of the most important is less invasive sample collection that is acceptable mainly in case of newborns, small children and older people. Other big pluses are low requirements for storing, transportation, qualified personnel and minimization of transfer of blood borne diseases. The volume of sample is much lower in DBS what requires more sensitive techniques for analysis. Also the stability of analytes and the influence of other components in blood matrix must be taken into account. This new technique of sampling was used for the first time in 1963 by Robert Guthrie in neonatal screening of phenylketonuria. From that time it has been used all over the world as a standard sampling method in newborn screening of different diseases. Due to a lot of advantages DBS becomes a very interesting sampling method also in other fields of analysis, for example in therapeutic drug monitoring, metabolomics studies, diagnostics of infectious diseases and so on. In our study we used this sampling method in analysis of fatty acids (FA), keto acids (KA), amino acids (AA) and glutathione (GSH). FA, after extraction from DBS with ethanol, were derivatized and their ethyl esters were separated by gas chromatography with flame ionization detector. KA were extracted with 1mmol/l HCl, AA and GSH were extracted with ethanol. All these analytes were separated in individual runs by reversed-phase high performance liquid chromatography (RP-HPLC) as 2-quinoline derivatives (KA), 1-cyano-2-substituted benz[f]isoindole derivatives (AA) or tricyclic isoindole derivatives (GSH) and monitored with fluorescence detector. Analytical parameters of method were satisfactory for AA, KA and GSH. In case of FA some problems with precision of measurement of low concentration FA were observed. Further research proceeds in this field. dried blood spot; amino acids; keto acids; fatty acids; glutathione; chromatography

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