title: [Simultaneous determination of guanidinoacetate, creatine and creatinine by liquid chromatography-tandem mass spectrometry: a diagnostic tool for creatine deficiency syndromes in body fluids and a perspective use on cultured fibroblasts].
abstract: Guanidinoacetate (GAA) and creatine (Cr) are creatine deficiency syndromes (CDS) biochemical markers. We describe a liquid chromatography - tandem mass spectrometry method (LC/MSMS) performing simultaneous analysis of GAA, Cr and creatinine (Crn). Study of Cr uptake by fibroblasts for Cr transporter defect diagnosis is also assessed. The three butylated compounds were separated by liquid chromatography and MSMS quantification was achieved by isotopic dilution with electrospray positive ion mode. Linearity was demonstrated from 0 to 600, 675 and 4500 μmol/L and limit of quantification was 0.1, 0.04 and 0.9 μmol/L for GAA, Cr, and Crn respectively. Intra- and inter-assay precision for each analyte was better than 11%, and standard recoveries ranged from 83 to 109%. Reference values in cerebrospinal fluid samples for subjects ≥14 years were also established for GAA and Cr. Five fibroblast cell lines were used for Cr uptake study. Cr uptake by fibroblasts increased with the Cr media concentrations and was significantly inhibited by 3-guanidinopropionate (500 μmol/L), a Cr transporter inhibitor (96h incubation, [Cr media] = 25 μmol/L, p<0.05). A reliable LC/MSMS method for the diagnosis of CDS was developed in different biological fluids. Finally, results of the Cr uptake study reinforce the interest of this technique to diagnose Cr transporter deficiencies.
authors.affiliation: Département de biochimie, toxicologie et pharmacologie, CHU, Hôpital A. Michalon, Grenoble, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Van Noolen
authors.forename: Denis
authors.initials: D
authors.lastname: Monneret
authors.forename: Véronique
authors.initials: V
authors.lastname: Ducros
authors.forename: Christelle
authors.initials: C
authors.lastname: Corne
authors.forename: Joël
authors.initials: J
authors.lastname: Lunardi
authors.forename: Patrice
authors.initials: P
authors.lastname: Faure
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000032:analysis / Q000134:cerebrospinal fluid; D001826:Body Fluids / Q000737:chemistry*; D020739:Brain Diseases, Metabolic, Inborn / Q000175:diagnosis; D018929:Cell Culture Techniques; D002460:Cell Line; D002853:Chromatography, Liquid / Q000379:methods*; D003401:Creatine / Q000032:analysis* / Q000134:cerebrospinal fluid / Q000172:deficiency; D003404:Creatinine / Q000032:analysis* / Q000134:cerebrospinal fluid; D003470:Culture Media; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005998:Glycine / Q000031:analogs & derivatives* / Q000032:analysis / Q000134:cerebrospinal fluid; D006146:Guanidines / Q000494:pharmacology; D006801:Humans; D008297:Male; D008875:Middle Aged; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors; D050480:Plasma Membrane Neurotransmitter Transport Proteins / Q000037:antagonists & inhibitors / Q000172:deficiency; D011422:Propionates / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
chemical_list: D015415:Biomarkers; D003470:Culture Media; D006146:Guanidines; D009419:Nerve Tissue Proteins; D050480:Plasma Membrane Neurotransmitter Transport Proteins; D011422:Propionates; C094770:SLC6A8 protein, human; D003404:Creatinine; C004946:glycocyamine; D003401:Creatine; D005998:Glycine; C005775:guanidinopropionic acid
keywords: creatine; creatine transporter; fibroblast; guanidinoacetate; tandem mass spectrometry