高效液相色谱-紫外可见检测器检测大鼠肝组织中的还原型谷胱甘肽  

Determination of reduced glutathione in rat livers by HPLC-UVD

作　　者：郗德凤[1] 王希峰[1] 苏冠民[1] 李琳[1] 杨燕[1] 邵丽华[1] 

机构地区：[1]山东大学公共卫生学院卫生检验研究所,济南250012

出　　处：《山东大学学报（医学版）》2010年第12期142-145,共4页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金资助项目(Y2007C079)

摘　　要：目的建立一种高效液相色谱-紫外可见检测器测定大鼠肝组织中还原型谷胱甘肽含量的方法。方法肝组织在冰浴条件下匀浆后经乙腈沉淀蛋白,离心,取上清液在硼酸缓冲液中与荧光剂4-氟-7-硝基-2,1,3-苯并氧杂恶二唑(NBD-F)进行衍生化反应。流动相为乙腈-0.02mol/L磷酸二氢钾溶液(23∶77,V/V),流速1.0mL/min,检测波长470nm,进样量10μL;采用外标法定量。结果还原型谷胱甘肽在0.200～125μg/mL范围内线性关系良好,相关系数为0.9999(n=5),最低检测限为0.0400μg/mL(S/N=3);样品加标回收率为96.2%～104.5%;日内相对标准偏差为1.1%～4.6%,日间标准偏差为1.9%～7.5%。结论本方法操作简便,快速,准确,灵敏度高,稳定性好,适合肝组织中还原型谷胱甘肽的测定。Objective To establish a high performance liquid chromatography with UV-absorbance detection method for determination of the level of reduced glutathione in rat livers.Methods After the liver tissue homogenate was made under the condition of ice bathing and tissue proteins precipitated by acetonitrile,the derivative reaction of the supernatant and fluorescent 4-fluoro-7-nitrobenzofuraza was carried out in borate buffer.The mobile phase was acetonitrile-0.02 mol/L potassium dihydrogen phosphate（23∶77,V/V）,the flow rate was 1.0 mL/min,the detection wave-length was 470 nm and the injection volume was 10 μL.The reaction mixture was quantified with the external standard method.Results The linear range of reduced glutathione was 0.200-125 μg/mL（r=0.999 9,n=5）,and the lowest limit of determination was 0.040 0 μg/mL（S/N=3）.The recovery rate was 96.2%-104.5%,the intra-day and inter-day relative standard deviations were 1.1%-4.6% and 1.9%-7.5%,respectively.Conclusion This method is simple,rapid,accurate,sensitive and stable,which is suitable for determination of reduced glutathione in the liver.

关 键 词：还原型谷胱甘肽 4-氟-7-硝基-2 1 3-苯并氧杂恶二唑 肝组织 高效液相色谱法 

分 类 号：R115[医药卫生—公共卫生与预防医学]