Quantitative determination of guanidinoacetate and creatine in dried blood spot by flow injection analysis-electrospray tandem mass spectrometry
| Autor: | Roberta Battini, Vincenzo Leuzzi, Italo Antonozzi, Teresa Giovanniello, Silvia Santagata, Cristiana Artiola, Claudia Carducci, Carla Carducci |
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| Rok vydání: | 2005 |
| Předmět: |
Amidinotransferases
Spectrometry Mass Electrospray Ionization guanidinoacetate Clinical Biochemistry Glycine gamt Ion suppression in liquid chromatography–mass spectrometry Tandem mass spectrometry Creatine Biochemistry chemistry.chemical_compound Reference Values tandem mass spectrometry Humans Child agat Chromatography High Pressure Liquid Detection limit Flow injection analysis Chromatography Biochemistry (medical) Selected reaction monitoring Infant Newborn nutritional and metabolic diseases Infant Reproducibility of Results General Medicine dried blood spot Dried blood spot Guanidinoacetate N-methyltransferase chemistry creatine Child Preschool Guanidinoacetate N-Methyltransferase Nervous System Diseases |
| Zdroj: | Clinica chimica acta; international journal of clinical chemistry. 364(1-2) |
| ISSN: | 0009-8981 |
| Popis: | Background Guanidinoacetate (GAA) and creatine (Cr) are reliable biochemical markers of primary creatine disorders. The aim of this study was to develop and validate a method for the determination of GAA and Cr in dried blood spot through the use of stable isotope dilution and flow injection analysis (FIA)-ESI-MS/MS. Methods Dried blood spots were extracted using methanol–water solution containing D3-Cr. After evaporation and formation of butyl esters, samples were analyzed using multiple reaction monitoring mode ( m / z 174.2→101.1 for GAA, 188.3→90.1 for Cr and 191.3→93.1 for D3-Cr). Results The analysis was very fast (1 min). The detection limits were 0.34 μmol/l of blood and 0.30 μmol/l of blood for Cr and GAA, respectively, and the response was linear over the range 0.25–12.5 μmol/l of blood for GAA and 3.57–624.7 μmol/l of blood for Cr. Recovery range was 93–101% for Cr and 94–105% for GAA and between-run CVs were 5.3% for GAA and 4.5% for Cr. Ion suppression effect was also studied. The method was applied to spots obtained from two patients affected by GAMT deficiency, four patients affected by AGAT deficiency (including a newborn) as well as 282 healthy subjects. Conclusions The detection of GAA in dried blood spot by FIA-ESI-MS/MS is a highly reliable and high throughput method for the diagnosis of GAMT and AGAT deficiencies and a possible tool for newborn screening of both these tractable disorders. |
| Databáze: | OpenAIRE |
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