An improved capillary electrophoresis method for measuring tissue metabolites associated with cellular energy state

被引:34
作者
Casey, TM [1 ]
Dufall, KG [1 ]
Arthur, PG [1 ]
机构
[1] Univ Western Australia, Dept Biochem, Nedlands, WA 6907, Australia
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1999年 / 261卷 / 03期
关键词
capillary electrophoresis; intracellular energy status; purine metabolism;
D O I
10.1046/j.1432-1327.1999.00324.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 reproductibility. 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 mu M 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.
引用
收藏
页码:740 / 745
页数:6
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