色谱

色谱 ›› 2013, Vol. 31 ›› Issue (11): 1064-1070.DOI: 10.3724/SP.J.1123.2013.04040

• 研究论文 • 上一篇    下一篇

可视化移动反应界面滴定法测定豆浆中蛋白质的含量

郭成叶1,2, 王后禹2, 张雷1, 樊柳荫2, 曹成喜2   

  1. 1. 上海师范大学生命与环境科学学院, 上海 200234;
    2. 上海交通大学生命科学技术学院, 微生物代谢国家重点实验室, 上海 200240
  • 收稿日期:2013-04-23 修回日期:2013-05-26 出版日期:2013-11-28 发布日期:2013-12-05
  • 通讯作者: 张雷, 曹成喜
  • 基金资助:

    科技部重大仪器专项基金项目(2011YQ030139);国家自然科学基金项目(21275099,21035004).

Determination of total protein content in soya-bean milk via visual moving reaction boundary titration

GUO Chengye1,2, WANG Houyu2, ZHANG Lei1, FAN Liuyin2, CAO Chengxi2   

  1. 1. College of Life and Environmental Sciences, Shanghai Normal University, Shanghai 200234, China;
    2. School of Life Sciences and Biotechnology of Shanghai Jiao Tong University, State Key Laboratory of Microbial Metabolism, Shanghai 200240, China
  • Received:2013-04-23 Revised:2013-05-26 Online:2013-11-28 Published:2013-12-05

PDF

203

摘要：

利用移动反应界面滴定(moving reaction boundary titration, MRBT)技术可视化快速准确测定豆浆中的总蛋白质含量。在MRBT中,移动反应界面(moving reaction boundary, MRB)是由阴极液中的氢氧根离子和固定在聚丙烯酰胺凝胶(polyacrylamide gel, PAG)中的豆浆蛋白质分子形成,通过酸碱指示剂来指示MRB的移动。利用MRB移动速度(VMRB)与蛋白质浓度存在的函数关系建立标准曲线。配对t检验MRBT测定结果与凯氏定氮相比在95%的置信区间内无显著性差异。MRBT测定所需时间为10 min左右,线性范围为2.0~14.0 g/L,检出限为0.05 g/L,日内相对标准偏差小于1.90%,日间相对标准偏差小于4.39%,回收率为97.41%~99.91%。此外,三聚氰胺等非蛋白质氮(non-protein nitrogen, NPN)的添加,对豆浆中蛋白质的测定结果影响较小。

关键词: 滴定, 电泳, 豆浆, 移动反应界面, 总蛋白质含量

Abstract:

A visual, rapid and accurate moving reaction boundary titration (MRBT) method was used for the determination of the total protein in soya-bean milk. During the process, moving reaction boundary (MRB) was formed by hydroxyl ions in the catholyte and soya-bean milk proteins immobilized in polyacrylamide gel (PAG), and an acid-base indicator was used to denote the boundary motion. The velocity of MRB has a relationship with protein concentration, which was used to obtain a standard curve. By paired t-test, there was no significant difference of the protein content between MRBT and Kjeldahl method at 95% confidence interval. The procedure of MRBT method required about 10 min, and it had linearity in the range of 2.0-14.0 g/L, low limit of detection (0.05 g/L), good precision (RSD of intra-day<1.90% and inter-day<4.39%), and high recoveries (97.41%-99.91%). In addition, non-protein nitrogen (NPN) such as melamine added into the soya-bean milk had weak influence on MRBT results.

Key words: electrophoresis, moving reaction boundary, soya-bean milk, titration, total protein content

中图分类号: