کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1969042 1538870 2015 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Screening for adenylosuccinate lyase deficiency using tandem mass spectrometry analysis of succinylpurines in neonatal dried blood spots
ترجمه فارسی عنوان
غربالگری کمبود آدنیلوسوکتینات لیاز با استفاده از تجزیه و تحلیل اسپکترومتری توده دوطرفه سوکسینولپورین ها در لکه های خون خشک شده نوزادان
کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
چکیده انگلیسی


• The preparation of isotopically labelled standards of succinylpurines is presented.
• LC–MS/MS method for determination of succinylpurines in neonatal DBS was developed.
• Patients with ADSL deficiency have elevated levels of succinylpurines in DBS.
• The new screening method may be used for retrospective diagnosis of ADSL deficiency.

ObjectivesStable 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 methodsSAICAr 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-13C4 and SAICAr-13C4, which were prepared via ADSL-catalysed reactions of fumarate-13C4 with adenosine monophosphate and aminoimidazole carboxamide ribotide, respectively, and subsequent alkaline phosphatase-catalysed dephosphorylation of the resulting products.ResultsThe detection of SAICAr and SAdo in DBS was linear over the range of 0–25 μmol/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, 0.03–4.7 μmol/L; SAdo, 1.5–21.3 μmol/L; n = 5) compared to those of the control subjects (SAICAr, 0–0.026 μmol/L; SAdo, 0.06–0.14 μmol/L; n = 31), even after DBSs from dADSL patients were stored for 2–23 years.ConclusionsWe 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.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Clinical Biochemistry - Volume 48, Issues 1–2, January 2015, Pages 2–7
نویسندگان
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