title: An improved capillary electrophoresis method for measuring tissue metabolites associated with cellular energy state.
abstract: An improved method for the measurement of tissue metabolites associated with cellular energetic state by capillary electrophoresis is described. This method allows 17 compounds present in a mixture of standards to be determined simultaneously within 43 min with good reproducibility. ATP, ADP, AMP, UTP, IMP, inosine, hypoxanthine, creatine, phosphocreatine, UDP-galactose, NAD and NADH were detected in samples of either rat heart tissue or rat neonatal cardiomyocytes. This method can detect compounds at concentrations of 5 microm in samples. Recoveries for ATP and phosphocreatine added to cardiomyocyte samples were 99.4 +/- 2.1% and 103.1 +/- 3.3%, respectively (mean +/- SEM, n = 3). Our method has been comprehensively validated and is capable of measuring a wider range of tissue metabolites important in assessing cellular energy status than existing methods.
authors.affiliation: Department of Biochemistry, The University of Western Australia, Nedlands 6907, Australia. tmcasey@cyllene.uwa.edu.au
authors.forename: T M
authors.initials: TM
authors.lastname: Casey
authors.forename: K G
authors.initials: KG
authors.lastname: Dufall
authors.forename: P G
authors.initials: PG
authors.lastname: Arthur
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002021:Buffers; D002138:Calibration; D015687:Cell Hypoxia; D002478:Cells, Cultured; D019075:Electrophoresis, Capillary / Q000379:methods*; D004734:Energy Metabolism; D006863:Hydrogen-Ion Concentration; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D010725:Phosphocreatine / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results
chemical_list: D002021:Buffers; D010725:Phosphocreatine; D009243:NAD; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D010100:Oxygen