Simultaneous determination of creatine and guanidinoacetate in plasma by liquid chromatography-tandem mass spectrometry (LC-MS/MS)

Sara Boenzi, Cristiano Rizzo, Vincenzo Maria Di Ciommo, Diego Martinelli, Bianca Maria Goffredo, Giancarlo La Marca, Carlo Dionisi-Vici

Research output: Contribution to journalArticle

Abstract

Guanidinoacetate (GAA) and creatine are reliable biochemical markers for primary and secondary creatine defects. We describe a method by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for simultaneous determination of plasma GAA and creatine. We analyzed 283 healthy subjects from 0 to 63 years old to obtain age-related control values. Methods: Plasma samples were extracted with acetonitrile containing 13C2-GAA and d3-creatine. Samples were analyzed by LC-MS/MS in positive ionisation mode, after derivatization to butyl-esters. Optimal chromatographic separation was achieved using a column Supelcosil™ LC-4.6. mm with isocratic elution in 5. min. Results: Run time was 5. min. Standard curves were linear from 0.05 to 200 μmol/L for creatine and from 0.02 to 40 μmol/L for GAA. Limit of detection (LOD) and limit of quantitation (LOQ) were respectively 0.005 and 0.05 μmol/L for creatine; LOD and LOQ were 0.002 and 0.02 μmol/L respectively for GAA. Intra and inter-assay CVs for creatine and GAA were 20 years old. Conclusion: A rapid and high sensitive LC-MS/MS method was developed and validated for determination of creatine and GAA in plasma and it could also be applied to other biological materials, such as CFS and urines. This method is useful for diagnoses of primary and also for secondary creatine defects that may occur in inherited metabolic diseases in which precursors of creatine biosynthesis are involved.

Original languageEnglish
Pages (from-to)792-798
Number of pages7
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume56
Issue number4
DOIs
Publication statusPublished - Dec 5 2011

Keywords

  • Creatine
  • Guanidinoacetate
  • LC-MS/MS

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Pharmaceutical Science
  • Spectroscopy
  • Clinical Biochemistry

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