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

ABSTRACT

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 The heart tissue was withdrawn for homogenate.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.