Simultaneous assay of isotopic enrichment and concentration of guanidinoacetate and creatine by gas chromatography-mass spectrometry.
A gas chromatography-mass spectrometry (GC-MS) method for the simultaneous measurement of isotopic enrichment and concentration of guanidinoacetate (GAA) and creatine in plasma sample for kinetic studies is reported. The method, based on preparation of the bis(trifluoromethyl)pyrimidine methyl ester derivatives of GAA and creatine, is robust and sensitive. The lowest measurable m(1) and m(3) enrichment for GAA and creatine, respectively, was 0.3%. The calibration curves for measurements of concentration were linear over ranges of 0.5 to 250microM GAA and 2 to 500microM for creatine. The method was reliable for inter- and intraassay precision, accuracy, and linearity. The technique was applied in a healthy adult to determine the in vivo fractional synthesis rate of creatine using primed-constant rate infusion of [1-(13)C]glycine. It was found that isotopic enrichment of GAA reached a plateau by 30min of infusion of [1-(13)C]glycine, indicating either a small pool size or a rapid turnover rate (or both) of GAA. In contrast, the tracer appearance in creatine was slow (slope=0.00097), suggesting a large pool size and a slow rate of synthesis of creatine. This method can be used to estimate the rate of synthesis of creatine in vivo in human and animal studies.
Kasumov
Takhar
T
Department of Pathobiology, Lerner Research Institute, Cleveland Clinic, OH 44195, USA.
Gruca
Lourdes L
LL
Dasarathy
Srinivasan
S
Kalhan
Satish C
SC
D000328:Adult; D053000:Analytic Sample Preparation Methods / Q000379:methods*; D002138:Calibration; D002247:Carbon Isotopes / Q000097:blood; D003401:Creatine / Q000097:blood*; D005215:Fasting; D005217:Fat Emulsions, Intravenous / Q000008:administration & dosage; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D005998:Glycine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000097:blood / Q000378:metabolism; D006801:Humans; D007201:Indicator Dilution Techniques; D007202:Indicators and Reagents; D007262:Infusions, Intravenous; D008954:Models, Biological; D009587:Nitrogen Isotopes / Q000097:blood; D010422:Pentanones
D002247:Carbon Isotopes; D005217:Fat Emulsions, Intravenous; D007202:Indicators and Reagents; D009587:Nitrogen Isotopes; D010422:Pentanones; C517499:hexafluoro-acetylacetone; C004946:glycocyamine; D003401:Creatine; D005998:Glycine