Screening for adenylosuccinate lyase deficiency using tandem mass spectrometry analysis of succinylpurines in neonatal dried blood spots
Jazyk angličtina Země Spojené státy americké Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
25445730
DOI
10.1016/j.clinbiochem.2014.10.004
PII: S0009-9120(14)00731-0
Knihovny.cz E-zdroje
- Klíčová slova
- Adenylosuccinate lyase deficiency, DBS, Dried blood spots, LC–MS/MS, Purine metabolism, Screening, Tandem mass spectrometry,
- MeSH
- adenosin analogy a deriváty krev MeSH
- adenylsukcinátlyasa krev nedostatek MeSH
- aminoimidazolkarboxamid analogy a deriváty krev MeSH
- autistická porucha MeSH
- chromatografie kapalinová MeSH
- izotopy uhlíku MeSH
- lidé MeSH
- limita detekce MeSH
- novorozenec MeSH
- poruchy metabolismu purinů a pyrimidinů krev diagnóza MeSH
- referenční standardy MeSH
- ribonukleosidy krev MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- test suché kapky krve metody MeSH
- Check Tag
- lidé MeSH
- novorozenec MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- adenosin MeSH
- adenylsukcinátlyasa MeSH
- aminoimidazolkarboxamid MeSH
- izotopy uhlíku MeSH
- ribonukleosidy MeSH
- succinyladenosine MeSH Prohlížeč
- succinylaminoimidazole carboxamide riboside MeSH Prohlížeč
OBJECTIVES: Stable isotope dilution coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the sensitive method for screening for various inherited metabolic disorders using dried blood spots (DBSs). We present a method for LC-MS/MS determination of succinyladenosine (SAdo) and succinylaminoimidazole carboxamide riboside (SAICAr), biomarkers for adenylosuccinate lyase deficiency (dADSL), in DBS. DESIGN AND METHODS: SAICAr and SAdo were separated on a Symmetry-C18 column and detected using positive electrospray ionisation in selected reaction monitoring mode. The quantification was performed using the isotopically labelled internal standards SAdo-(13)C4 and SAICAr-(13)C4, which were prepared via ADSL-catalysed reactions of fumarate-(13)C4 with adenosine monophosphate and aminoimidazole carboxamide ribotide, respectively, and subsequent alkaline phosphatase-catalysed dephosphorylation of the resulting products. RESULTS: The detection of SAICAr and SAdo in DBS was linear over the range of 0-25μmol/L. The respective intra-assay and inter-assay imprecision values were less than 10.7% and 15.2% for SAICAr and 4.7% and 5.7% for SAdo. The recoveries from DBS spiked with different concentrations of SAICAr and SAdo were between 94% and 117%. The concentrations of SAICAr and SAdo were higher in the archived DBS from dADSL patients (SAICAr, 0.03-4.7μmol/L; SAdo, 1.5-21.3μmol/L; n=5) compared to those of the control subjects (SAICAr, 0-0.026μmol/L; SAdo, 0.06-0.14μmol/L; n=31), even after DBSs from dADSL patients were stored for 2-23years. CONCLUSIONS: We developed and validated a method of succinylpurine analysis in DBS that improves selective screening for dADSL in the paediatric population and may be used for retrospective diagnosis to aid the genetic counselling of affected families.
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