Simultaneous determination of myocardial creatine phosphate and adenine nucleotides by reversed-phase HPLC.
Simultaneous determination of myocardial creatine phosphate and adenine nucleotides by reversed-phase HPLC.
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DOI：
10.1002/bmc.1130020406
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
R. Sanduja;G. Ansari;P. Boor
中科院分区：
文献类型：
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作者：
R. Sanduja;G. Ansari;P. Boor
已开发了等度HPLC系统，该系统允许快速（单次运行20 min）测量冷冻夹持和冻干心肌组织提取物中的磷酸肌酸（PCr）和腺嘌呤核苷酸（ATP、ADP和AMP）。采用RP 18色谱柱和双波长分光光度计，在210和254 nm波长下进行分离。溶剂为30 mM磷酸二氢钾、15 mM四丁基硫酸氢铵（pH 6.7）、19%（v/v）乙腈。实现了这些高能化合物的明显分离（用标准样品的保留时间确认）。标准曲线呈线性。在离体大鼠心脏中，获得了以下值（mumol/g干重，平均值+/- SEM）：ATP 21.5 +/- 1.3，ADP 4.6 +/- 0.2，AMP 1.5 +/- 1.1和PCr 32.5 +/- 1.3;这与先前发表的该组织中高能化合物的值一致。
An isocratic HPLC system has been developed which allows for the rapid (single run of 20 min) measurement of creatine phosphate (PCr) and adenine nucleotides (ATP, ADP and AMP) in extracts from freeze-clamped and freeze-dried myocardial tissues. The separation was achieved at room temperature by using a RP18 column and a dual variable wavelength spectrophotometer, set at 210 and 254 nm. The solvent was 30 mM potassium dihydrogen phosphate, 15 mM tetrabutylammonium hydrogen sulfate, pH 6.7, 19% (v/v) acetonitrile. A distinct separation (confirmed with the retention time of standard sample) of these high energy compounds was achieved. Standard curves were linear. In isolated rat hearts the following values were obtained (mumol/g dry wt, mean +/- SEM): ATP 21.5 +/- 1.3, ADP 4.6 +/- 0.2, AMP 1.5 +/- 1.1 and PCr 32.5 +/- 1.3; which are consistent with previously published values for high energy compounds in this tissue.