An improved capillary electrophoresis method for measuring tissue metabolites associated with cellular energy state.
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.
Casey
T M
TM
Department of Biochemistry, The University of Western Australia, Nedlands 6907, Australia. tmcasey@cyllene.uwa.edu.au
Dufall
K G
KG
Arthur
P G
PG
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
D002021:Buffers; D010725:Phosphocreatine; D009243:NAD; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D010100:Oxygen