反相离子对高效液相色谱法测定心肌组织中的三磷酸腺苷

Determination of the Adenosine Triphosphate in Myocardial Tissue by Ion-Pair Reversed-Phase High Performance Liquid Chromatography

采用反相离子对高效液相色谱法测定心肌组织中三磷酸腺苷 (ATP)的含量。样品经高氯酸溶液沉淀蛋白 ,上清液用KOH溶液中和后用反相离子对高效液相色谱法分离测定。色谱柱为SpherisorbODS2柱 ,流动相为甲醇 KH2 PO4缓冲液 (内含 5mmol/LIPR A离子对试剂 ) ,在 2 59nm波长处检测。方法最低检测限为 2mg/L,在 5mg/L～ 1 0 0 mg/L范围内有良好的线性关系 (r=0 9998) ,方法的回收率为 97 8%～ 1 0 4 % ,日内精密度 <4 85% ,日间精密度 <8 81 %。方法准确、灵敏、快速 ,适用于动物和人心肌组织中ATP含量的测定。

ATP depletion is an important factor contributing to the incomplete recovery of ventricular function following ischemia and reperfusion A simple and rapid method for determination of adenosine triphosphate(ATP) in myocardial tissue by RP HPLC was developed The chromatographic conditions were as follows:Spherisorb ODS2 column, V(40 mmol/L KH\-2PO\-4 and 5 mmol/L TBAP )∶V(methanol)=74∶26 (pH 6 2) mobile phase and UV detector at 259 nm The ATP in myocardium was extracted with 0 4 mol/L HClO\-4 The calibration curve showed a good linearity in 5 mg/L 100 mg/L(r=\{0 999 8\}) The recovery ranged from 97 8% to 104 1% The limit of detection was 2 mg/L The verified results demonstrated that this method is precise, accurate and can be used for determination of ATP in myocardial tissue