Analysis of aminoimidazole ribosides by capillary electrophoresis — Diagnosing defects in second part of purine biosynthetic pathway

Only three inherited metabolic defects have been identified in purine de novo synthesis (PDNS). We present here CE methods for diagnosing defects in the second half of PDNS (from sixth to tenth enzymatic conversion) based on analysis of aminoimidazole ribosides – dephosphorylated intermediates – in...

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Bibliographic Details
Published in:Clinica chimica acta 2007-02, Vol.376 (1-2), p.184-189
Main Authors: Hornik, Petr, Vyskočilová, Petra, Friedecký, David, Janošťáková, Anna, Adamová, Kateřina, Adam, Tomáš
Format: Article
Language:English
Subjects:
Adenylosuccinate Lyase - deficiency
Adolescent
Amino Acid Metabolism, Inborn Errors - diagnosis
Animals
Biosynthetic Pathways
Capillary electrophoresis
Cell Line
Child
Child, Preschool
CHO Cells
Cricetinae
Cricetulus
Electrophoresis, Capillary - methods
Female
Humans
Imidazoles - urine
Inborn diseases
Infant
Male
Molecular Structure
Purine Nucleosides - biosynthesis
Purines
Reference Values
Time Factors
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Summary:Only three inherited metabolic defects have been identified in purine de novo synthesis (PDNS). We present here CE methods for diagnosing defects in the second half of PDNS (from sixth to tenth enzymatic conversion) based on analysis of aminoimidazole ribosides – dephosphorylated intermediates – in urine. Assays were performed in an uncoated fused-silica capillary using two electrophoretic separation systems: 60 mmol/l borate — 2-amino-2-methyl-1-propanol–80 mmol/l sodium dodecylsulfate (pH 9.6) and 200 mmol/l phosphate — sodium (pH 1.8). The reported conditions allowed separation of all metabolites from major urinary constituents with analysis time less than 10 min and separation efficiency of 220 and 350 thousands theoretical plates per meter for borate and phosphate system, respectively. The intra- and interday imprecisions were less than 4.4% and 9.9% CV. Potential usefulness of the methods was demonstrated on samples from a patient with adenylosuccinate lyase deficiency and Chinese hamster ovary cell lines defective in PDNS. CE is a useful and effective tool in the analysis of aminoimidazole ribosides which enables diagnosis of known as well as not so far identified inherited defects of PDNS pathway.
ISSN:0009-8981
1873-3492
DOI:10.1016/j.cca.2006.08.020