HPLC法测定心衰大鼠心肌中ATP、ADP与AMP的含量

Determination of ATP,ADP and AMP in Cardiac Muscle of Rats with Heart Failure by HPLC

目的：建立大鼠心肌组织ATP、ADP、AMP 的含量测定方法,考察心衰大鼠的ATP、ADP、AMP定量变化。方法：色谱柱为Thermo Hypersil C18柱（250 mm ×4.6 mm,5μm）；流动相为6.8 g·L-1磷酸二氢钾和1.164 g·L-1氢氧化钠溶液；检测波长为254 nm。 SD大鼠随机分为对照组和试验组,分别腹腔注射生理盐水和多柔比星,给药6周后取心脏组织制备匀浆,用HPLC法测定大鼠心脏组织中ATP、ADP和AMP的含量。结果：ATP、ADP和AMP在0.625~40μg·ml-1呈良好的线性（ r〉0.99）,最低检测限和最低定量限为0.625μg·ml-1,精密度RSD均〈9.50%,回收率为93.20%~108.0%,稳定性RSD 为3.0%~14.0%。多柔比星组ATP的含量均明显低于生理盐水组,而AMP和ADP含量则明显高于生理盐水组（P〈0.01）。结论：HPLC法测定心肌组织中ATP、ADP和AMP含量满足生物样本的测定要求,可用于考察蒽环类抗肿瘤药物对心衰大鼠的影响。

Objective：To establish the determination method for ATP, ADP and AMP in the myocardial tissue of rats and research the changes of ATP, ADP and AMP in heart fallure rats. Methods： A Thermo Hypersil C18 （250 mm × 4. 6 mm,5 μm） column was used. The solution of 0. 68% potassium dihydrogen phosphate and 0. 116% soduim hydroxide was applied as the mobilie phase; The detection wavelength was at 254 nm. The SD rats randomly divided into 2 groups received saline and adriamycin by intraperitoneal in-jection for 6 weeks, respectively. The heart tissue was withdrawn for homogenate. The contents of ATP, ADP and AMP in rat heart tis-sue were determined by HPLC. Results：The calibration curves were linear from 0. 625μg·ml-1 to 40μg·ml-1 for ATP, ADP and AMP. The limit of detection and limit of quantification were 0. 625μg·ml-1（r〉0. 99）. The precision of RSD was less than 9. 50%. The accuracy was 93. 2%-108. 0%. The method stability （RSD） was from 3. 0% to 14. 0%. The content of ATP in heart fallure mod-el group induced by adriamycin was significantly lower than that in saline group,while that of ADP were much higher than that in saline group（P〈0. 01） and AMP. Conclusion：The HPLC method for the content determination of ATP, ADP and AMP in myocardial tissue meets the requirements of the determination of biological samples, which can be used to study the effects of anthracycline-based antitu-mor drugs in heart fallure rats.