食品科学

• 分析检测 • 上一篇    下一篇

酶联免疫吸附结合荧光分析法测定乳粉加速贮藏过程中的褐变产物

刘 玲,杨双华,纪淑娟,Leif Horsfelt SKIBSTED   

  1. 1.沈阳农业大学食品学院,辽宁 沈阳 110866;2.哥本哈根大学生命学部食品系,哥本哈根 2200
  • 出版日期:2013-09-25 发布日期:2013-09-27

Determination ELISA Coupled with Fluorescence Spectroscopy of Browning Products of Milk Powder during Accelerated Storage

LIU Ling,YANG Shuang-hua,JI Shu-juan,Leif Horsfelt SKIBSTED   

  1. 1. College of Food Science, Shenyang Agricultural University, Shenyang 110866, China;
    2. Department of Food Science, Faculty of Life Sciences, University of Copenhagen, Copenhagen 2200, Denmark
  • Online:2013-09-25 Published:2013-09-27

摘要:

在65℃封闭的加速贮藏条件下,对3种不同类型乳粉(全脂乳粉、脱脂乳粉和酪乳粉)的褐变产物——末端糖基化产物(AGEs)进行研究和检测。在确立酶联免疫吸附(ELISA)法测定末端糖基产物含量的基础上,结合荧光分析测定乳粉中蛋白末端糖基化所产生的荧光物质。结果表明:AGEs产生的荧光物质和AGEs含量随着贮藏时间的延长而增加。酶联免疫吸附法采用初级抗体和酶标二抗稀释倍数均为2500倍时可以有效测定褐变乳粉中AGEs含量,其中加速贮藏20d后乳粉中AGEs含量最高,可达到0.61ng/mL。最低检出质量浓度达到0.006ng/mL,最大误差率在5%左右,重复性良好。因此,乳粉褐变的荧光强度和AGEs含量的ELISA检测可以作为评价乳粉质量的有效方法。

关键词: 乳粉, 非酶褐变, 末端糖基化产物, 酶联免疫吸附法, 荧光分析

Abstract:

This study aimed to determine the browning products—advanced glycation end products (AGEs) in milk powder
stored under accelerated conditions at 65 ℃ for 20 days by ELISA coupled with fluorescence spectroscopy. Results indicated
that higher amounts of AGEs and fluorescent compounds were formed with prolonged storage time. ELISA could effectively
determine the content of AGEs in milk powder when the concentration of both primary antibody and HRP-labeled secondary
antibody was diluted to 2500 times. The highest content of AGEs in whole milk powder was observed, reaching 0.61 ng/mL,
after 20 days of storage, and the minimum detected concentration was 0.006 g/mL. The maximum relative error was around
5% and good repeatability was found. Therefore, fluorescence intensity and the level of AGEs could be applied to evaluate
the quality of milk powder.

Key words: milk powder, browning products, advanced glycation products, enzyme-linked immunosorbent assay(ELISA), fluorescence analysis