高效液相色谱-电化学法检测束缚挤压伤大鼠血清儿茶酚胺含量变化  

Determination of serum catecholamine in rats with restraint and crush injury by HPLC-ECD

作　　者：于梦洋 吕琪[1] 王鑫[1] 丁辉[1] 刘子泉[1] 曹娟[1] 刘晋阳[1] 王雪[1] 樊毫军[1] 侯世科[1] 

机构地区：[1]武警后勤学院附属医院救援研究所,天津300162

出　　处：《武警后勤学院学报（医学版）》2016年第5期345-349,共5页Journal of Logistics University of PAP（Medical Sciences）

基　　金：国家自然科学基金资助项目(71473265;71533008);武警后勤学院基础研究项目(WHJ2015014)

摘　　要：【目的】建立高效液相色谱-电化学法检测儿茶酚胺的方法,并检测束缚挤压伤大鼠血清儿茶酚胺的变化。【方法】制作束缚挤压伤大鼠模型,分别留取造模0、6、12、24 h大鼠血清样本并进行处理;应用高效液相检测仪和电化学检测仪对大鼠血清中去甲肾上腺素(NE)、肾上腺素(E)、多巴胺(DA)含量进行测量。【结果】肾上腺素、去甲肾上腺素及多巴胺的3个峰分离效果较好,峰形对称性较好,无内源性杂质干扰峰,各物质的回收率和RSD值(100%)分别为:去甲肾上腺素:98.54%、1.85%;肾上腺素:99.21%、2.36%;多巴胺:97.31%、2.42%。血清NE浓度在挤压后12 h明显升高(与0 h相比,P<0.05),血清E和DA浓度在挤压后6 h即开始明显升高(与0 h相比,P<0.05),随后NE、E、DA浓度不同程度减低后再次升高,NE和DA浓度在12 h达到峰值后回落,E浓度持续升高。【结论】确立了简便、快捷、稳定的高效液相色谱-电化学检测儿茶酚胺的方法,验证了束缚挤压伤大鼠早期存在明显应激反应。【Objective】To establish a method of high performance liquid chromatography with electrochemical detection（HPLC-ECD）to detect the changes of serum catecholamine in rats with restraint and crush injury.【Methods】The rat models with restraint and crush injury were made. Their serum samples were collected and treated respectively at 0 h, 6 h, 12 h, 24 h. Then the levels of norepinephrine（NE）, epinephrine（E） and dopamine（DA） in rat serum were determined with high performance liquid chromatography detector and electrochemical detection.【Results】The separation effect of E, NE and DA peaks was ideal and the peak shape was symmetrical without the interference of endogenous impurity. The recovery rates and RSD values were as bellow： NE： 98.54%, 1.85%; E： 99.21%, 2.36%;DA： 97.31%, 2.42%. Serum concentration of NE increased significantly in 12 h group（vs. 0 h group, P〈0.05）, while serum concentrations of E and DA increased significantly in 6 h group（vs. 0 h group, P〈0.05）. Subsequently, the concentrations of NE, E and DA decreased in different degrees and increased again. The concentrations of NE and DA reached the peak at 12 th h and fell back, while the concentration of E rose continuously.【Conclusion】A simple, convenient and stable method of HPLC-ECD is established to determine the changes of catecholamine. It is verified that rats have obvious stress reactions in the early stage of restraint and crush injury.

关 键 词：高效液相色谱法 挤压伤 束缚 儿茶酚胺 

分 类 号：Q503[生物学—生物化学]