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Screening for adenylosuccinate lyase deficiency using tandem mass spectrometry analysis of succinylpurines in neonatal dried blood spots.
Clin Biochem 2015; 48(1-2):2-7CB

Abstract

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.

Authors+Show Affiliations

Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, 128 08 Praha 2, Czech Republic. Electronic address: marie.zikanova@lf1.cuni.cz.Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, 128 08 Praha 2, Czech Republic. Electronic address: jkrijt@lf1.cuni.cz.Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, 128 08 Praha 2, Czech Republic. Electronic address: vaclava.skopova@lf1.cuni.cz.Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, 128 08 Praha 2, Czech Republic. Electronic address: Matyas.Krijt@staff.cuni.cz.Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, 128 08 Praha 2, Czech Republic. Electronic address: veronika.baresova@lf1.cuni.cz.Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, 128 08 Praha 2, Czech Republic. Electronic address: skmoch@lf1.cuni.cz.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25445730

Citation

Zikanova, Marie, et al. "Screening for Adenylosuccinate Lyase Deficiency Using Tandem Mass Spectrometry Analysis of Succinylpurines in Neonatal Dried Blood Spots." Clinical Biochemistry, vol. 48, no. 1-2, 2015, pp. 2-7.
Zikanova M, Krijt J, Skopova V, et al. Screening for adenylosuccinate lyase deficiency using tandem mass spectrometry analysis of succinylpurines in neonatal dried blood spots. Clin Biochem. 2015;48(1-2):2-7.
Zikanova, M., Krijt, J., Skopova, V., Krijt, M., Baresova, V., & Kmoch, S. (2015). Screening for adenylosuccinate lyase deficiency using tandem mass spectrometry analysis of succinylpurines in neonatal dried blood spots. Clinical Biochemistry, 48(1-2), pp. 2-7. doi:10.1016/j.clinbiochem.2014.10.004.
Zikanova M, et al. Screening for Adenylosuccinate Lyase Deficiency Using Tandem Mass Spectrometry Analysis of Succinylpurines in Neonatal Dried Blood Spots. Clin Biochem. 2015;48(1-2):2-7. PubMed PMID: 25445730.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Screening for adenylosuccinate lyase deficiency using tandem mass spectrometry analysis of succinylpurines in neonatal dried blood spots. AU - Zikanova,Marie, AU - Krijt,Jakub, AU - Skopova,Vaclava, AU - Krijt,Matyas, AU - Baresova,Veronika, AU - Kmoch,Stanislav, Y1 - 2014/10/23/ PY - 2014/09/03/received PY - 2014/10/14/revised PY - 2014/10/15/accepted PY - 2014/12/3/entrez PY - 2014/12/3/pubmed PY - 2015/9/9/medline KW - Adenylosuccinate lyase deficiency KW - DBS KW - Dried blood spots KW - LC–MS/MS KW - Purine metabolism KW - Screening KW - Tandem mass spectrometry SP - 2 EP - 7 JF - Clinical biochemistry JO - Clin. Biochem. VL - 48 IS - 1-2 N2 - 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. SN - 1873-2933 UR - https://www.unboundmedicine.com/medline/citation/25445730/Screening_for_adenylosuccinate_lyase_deficiency_using_tandem_mass_spectrometry_analysis_of_succinylpurines_in_neonatal_dried_blood_spots_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0009-9120(14)00731-0 DB - PRIME DP - Unbound Medicine ER -