食品科学

• 分析检测 • 上一篇    下一篇

DTNB比色法测定金属硫蛋白含量

吴云辉,王俊坤,孙继鹏,曾嘉玲   

  1. 1.厦门海洋职业技术学院,福建 厦门 361100;2.国家海洋局第三海洋研究所,福建 厦门 361005;
    3.中国海洋大学食品科学与工程学院,山东 青岛 266003
  • 出版日期:2013-08-25 发布日期:2013-09-03

Colormetric Determination of Metallothionein Using DTNB

WU Yun-hui,WANG Jun-kun,SUN Ji-peng,ZENG Jia-ling   

  1. 1. Xiamen Ocean Vocational College, Xiamen 361100, China;2. Third Institue of Oceanography State Oceanic Administration,Xiamen 361005, China;3. College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China
  • Online:2013-08-25 Published:2013-09-03

摘要:

采用5,5’-二硫硝基苯甲酸(DTNB)作为金属硫蛋白(MT)巯基衍生化试剂,通过优化反应条件,应用比色法测定412nm波长处显色产物硫代硝基苯甲酸阴离子(TNBA)的量确定金属硫蛋白含量。结果表明:MT含量在2~15μg和10~150μg范围内呈良好线性,线性回归方程分别为A412nm=0.0059C+0.0251和A412nm=0.0052C+0.0201,相关系数分别为R2=0.9942和R2=0.9997,平均回收率为99.94%(n=6,RSD=0.27%),精密度良好(n=6,RSD=0.7%),MT最低定量限可达到2μg,从而建立了一种操作简便易行的金属硫蛋白快速检测方法。

关键词: 金属硫蛋白, 5, 5&rsquo, -二硫硝基苯甲酸(DTNB), 测定

Abstract:

A corlometric method for the determination of metallothionein (MT) was established using 5,5’-dithio bis-
(2-nitrobenzoic acid) (DTNB) as the derivatizing agent and measuring the absorbance of the colored product TNBA
at 412 nm. The results showed that a linear relationship of MT was obtained within the concentration range of 2–15 μg
and 10–150 μg. The corresponding equations were A412 nm = 0.0059C + 0.0251 (R2 = 0.9942), and A412 nm = 0.0052C +
0.0201 (R2 = 0.9997). The average recovery was 99.94% (n = 6, RSD = 0.27%), with good precision (n = 6, RSD = 0. 7%).
The limit of detection of the method was 2 μg. In conclusion this study has developed a simple and rapid method to
determine MT.

Key words: metallothionein (MT);5,5&rsquo, -dithio bis-(2-nitrobenzoic acid) (DTNB);determination