高效液相色谱法测定慢性压力负荷性心衰大鼠心肌组织中腺苷酸的含量  

Determination of adenosine phosphates in myocardium of rats with pressure overload-induced chronic heart failure by HPLC

作　　者：杨娟[1] 黄敬群[1] 曹雪滨[2] 李红[1] 王艳飞[2] 刘红[1] 

机构地区：[1]中国人民解放军第252医院药剂科,河北保定071000 [2]中国人民解放军第252医院心内科,河北保定071000

出　　处：《中国医院药学杂志》2009年第1期11-14,共4页Chinese Journal of Hospital Pharmacy

基　　金：全军医学科学技术研究"十一五"计划课题资助项目(编号:2005064030)

摘　　要：目的:建立测定大鼠心肌组织中三磷酸腺苷、二磷酸腺苷、一磷酸腺苷(ATP、ADP、AMP)含量的高效液相色谱法,并观察慢性压力负荷性心衰大鼠心肌组织中腺苷酸含量的变化。方法:SD大鼠随机分为心衰模型组和假手术对照组,按腹主动脉缩窄法建立慢性压力负荷性心衰大鼠模型。色谱柱为VP-ODSC18柱(250mm×4.6mm,5μm);流动相为50mmol.L-1磷酸盐缓冲液(pH6.5);流速为1.0mL·min-1;柱温25℃;检测波长259nm;进样量20μL。结果:ATP、ADP、AMP的标准液与峰面积均呈良好的线性关系(r=0.9997～0.9999),平均回收率分别为96.30%,97.64%和103.28%。日内和日间精密度分别在1.15%和4.13%以下。心衰大鼠与假手术对照大鼠相比,ATP和ADP的含量明显下降,而AMP明显升高。结论:本法操作简便,准确快速,可以用于测定大鼠心肌组织中腺苷酸的含量。OBJECTIVE To develop a high performance liquid chromatography （HPLC） method to determine the content of ATP, ADP and AMP in myocardium of rats, and to observe the change of adenosine phosphates in chronic heart failure which resulted from the chronic excessive post-load. METHODS Male SD rats were divided into abdominal aortic-banded or shatmoperated groups. The model of rats with pressure overload-induced chronic heart failure were established by partially banding abdominal aorta. HPLC analysis was preformed using a VP-ODS C18 column （250 mm×4. 6 mm, 5μm）. The mobile phase consisted of 50 mmol.L^-1 tribasic potassium phosphate buffer solution （pH 6. 5）. The flow-rate was 1 mL·min^-1 and the temperature of column was about 25℃. The detection wavelength was 259 nm and the injection volume was 20 μL. RESULTS The calibration curve showed good linearity（r = 0. 999 7 - 0. 999 9）and the average recovery rates of ATP, ADP and AMP were 96. 30%, 97. 64% and 103.28%. The within-day and betweemday precisions were less than 1.15% and 4. 13%, respectively. The study showed that as compared with sham operated groups, ATP and ADP levels were significantly decreased in abdominal aortic-banded group, hut AMP levels were increased significantly. CONCLUSION This analytical method is simple, rapid and specific in detecting the content of adenosine phosphates in myocardium of rats.

关 键 词：高效液相色谱法 心肌 腺苷酸 心力衰竭 

分 类 号：R927.2[医药卫生—药学]