食品科学

• 分析检测 • 上一篇    下一篇

ELISA法检测牛奶中头孢氨苄残留

卞素敏,扈洪波,邢仕歌,张 悦,熊齐荣,钟 强,金 涌,储晓刚   

  1. 1.中国检验检疫科学研究院食品安全研究所,北京 100123;2.中国农业大学食品科学与营养工程学院,北京 100083
  • 出版日期:2013-10-25 发布日期:2013-09-28

Development of ELISA Method for Detection of Cepfalexin Residue in Milk

BIAN Su-min,HU Hong-bo,XING Shi-ge,ZHANG Yue,XIONG Qi-rong,ZHONG Qiang,JIN Yong,CHU Xiao-gang   

  1. 1. Institute of Food Safety, Chinese Academy of Inspection and Quarantine, Beijing 100123, China;
    2. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China
  • Online:2013-10-25 Published:2013-09-28

摘要:

应用头孢氨苄单克隆抗体,建立牛奶中头孢氨苄残留的间接竞争酶联免疫吸附测定(ELISA)检测方法,其牛奶中检测线性范围为1.0~50.0ng/mL,检测限为1.0ng/mL,半数抑制质量浓度为4.1ng/mL,牛奶的加标回收率分别为96.0%、98.2%及93.3%。头孢氨苄抗体与头孢拉定、头孢羟氨苄、头孢克洛及头孢克肟的交叉反应率分别为160.0%、80.0%、28.4%和3.7%。此外,一种简单、有效的牛奶纯化方法得以建立。对比ELISA和超高效液相色谱-串联质谱实验结果,显示两种方法有很好的数据相关性(R2=0.9988)。

关键词: 头孢氨苄, 单克隆抗体, 间接竞争酶联免疫吸附测定, 牛奶, 残留检测, 超高效液相色谱-串联质谱

Abstract:

Based on cafalexin monoclonal antibody, an indirect competitive enzyme-linked immunosorbent assay (ELISA)
to determine cefalexin residues in milk was established. The linear detection range of the ELISA method in milk was 1.0
to 50.0 ng/mL, with limit of detection of 1.0 ng/mL, and IC50 value of 4.1 ng/mL. The recovery rates for spiked milk with
50, 100 ng/mL and 200 ng/mL of cafalexin were 96.0%, 98.2% and 93.3%, respectively. Satisfactory cross-reactivity of the
antibody with cefradine, cefadroxil, cefaclor and cefixme was also observed (160.0%, 80.0%, 28.4% and 3.7%, respectively).
In addition, a simple and effective sample extraction method was established. The ELISA assay was validated by comparing
its results with those obtained by using UPLC-MS-MS, with a good correlation (R2) of 0.9988.

Key words: cefalexin, monoclonal antibody, indirect competitive ELISA, milk, residue detection, UPLC-MS-MS