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山东大学学报(医学版) ›› 2010, Vol. 48 ›› Issue (12): 142-145.

• 论文 • 上一篇    下一篇

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

郗德凤,王希峰,苏冠民,李琳,杨燕,邵丽华   

  1. 山东大学公共卫生学院卫生检验研究所, 济南 250012
  • 收稿日期:2010-07-27 发布日期:2010-12-16
  • 通讯作者: 邵丽华(1958- ),女,教授,主要从事色谱分析在卫生检验中的应用研究。 E-mail:slh@sdu.edu.cn
  • 作者简介:郗德凤(1985- ),女,硕士研究生,主要从事色谱分析在卫生检验中的应用研究。
  • 基金资助:

    山东省自然科学基金资助项目(Y2007C079)

Determination of reduced glutathione in the rat livers by HPLC-UVD

CHI De-feng, WANG Xi-feng, SU Guan-min, LI Lin, YANG Yan, SHAO Li-hua   

  1. Department of Sanitary Detection, School of Public Health, Shandong University, Jinan 250012, China
  • Received:2010-07-27 Published:2010-12-16

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。

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

Abstract:

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 precipitated tissue proteins 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-0.02mol/L potassium dihydrogen phosphate (23∶77, V/V), the flow rate was 1.0mL/min, the detection wave-length was 470nm and the injection volume was 10μL. The reaction mixture was quantified with the external standard method. Results     The linear range of reduced glutathione was 0.200-125μg/mL (r=0.9999, n=5), and the lowest limit of determination was 0.0400μg/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.

Key words: Reduced glutathione; 4-fluoro-7-nitrobenzofuraza; Liver tissue; High performance liquid chromatography

中图分类号: 

  • R115
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