title: Simultaneous determination of guanidinoacetate, creatine and creatinine in urine and plasma by un-derivatized liquid chromatography-tandem mass spectrometry.
abstract: BACKGROUND
  Creatine plays an important role in the storage and transmission of phosphate-bound energy. The cerebral creatine deficiency syndromes (CCDS) comprise three inherited defects in creatine biosynthesis and transport. They are characterized by mental retardation, speech and language delay and epilepsy. All three disorders cause low-creatine signal on brain magnetic resonance spectroscopy (MRS); however, MRS may not be readily available and even when it is, biochemical tests are required to determine the underlying disorder.
  
  
  METHODS
  Analysis was performed by liquid chromatography-tandem mass spectrometry in positive ionization mode. Samples were analysed underivatized using a rapid 'dilute and shoot' approach. Chromatographic separation of the three compounds was achieved. Stable isotope internal standards were used for quantification.
  
  
  RESULTS
  Creatine, creatinine and guanidinoacetate were measured with a 2.5 minute run time. For guanidinoacetate, the standard curve was linear to at least 5000 mumol/L and for creatine and creatinine it was linear to at least 25 mmol/L. The lower limit of quantitation was 0.4 mumol/L for creatine and guanidinoacetate and 0.8 mumol/L for creatinine. Recoveries ranged from 86% to 106% for the three analytes. Intra- and inter-assay variation for each analyte was <10% in both urine and plasma.
  
  
  CONCLUSIONS
  A tandem mass spectrometric method has been developed and validated for the underivatized determination of guanidinoacetate, creatine and creatinine in human urine and plasma. Minimal sample preparation coupled with a rapid run time make the method applicable to the routine screening of patients with suspected CCDS.
authors.affiliation: Biochemical Genetics Unit, Box 247, Addenbrookes Hospital, Hills Road, Cambridge CB2 0QQ, UK. rachel.carling@nhs.net
authors.forename: R S
authors.initials: RS
authors.lastname: Carling
authors.forename: S L
authors.initials: SL
authors.lastname: Hogg
authors.forename: T C
authors.initials: TC
authors.lastname: Wood
authors.forename: J
authors.initials: J
authors.lastname: Calvin
mesh_terms: D000328:Adult; D001774:Blood Chemical Analysis / Q000379:methods; D020739:Brain Diseases, Metabolic, Inborn / Q000097:blood / Q000175:diagnosis / Q000652:urine; D002648:Child; D002675:Child, Preschool; D002853:Chromatography, Liquid / Q000379:methods; D003401:Creatine / Q000097:blood* / Q000172:deficiency / Q000652:urine*; D003404:Creatinine / Q000097:blood* / Q000652:urine*; D005998:Glycine / Q000031:analogs & derivatives* / Q000097:blood / Q000652:urine; D050937:Guanidinoacetate N-Methyltransferase / Q000172:deficiency; D006801:Humans; D026901:Membrane Transport Proteins / Q000172:deficiency; D013577:Syndrome; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D055815:Young Adult
chemical_list: D026901:Membrane Transport Proteins; C080679:creatine transporter; D003404:Creatinine; C494506:GAMT protein, human; D050937:Guanidinoacetate N-Methyltransferase; C004946:glycocyamine; D003401:Creatine; D005998:Glycine