食品科学

• 工艺技术 • 上一篇    下一篇

纳米免疫磁珠富集猪肉中的盐酸克伦特罗

李超辉,熊勇华,郭 亮,吴科盛,梁语嫣   

  1. 1.南昌大学 食品科学与技术国家重点实验室,江西 南昌 330047;
    2.南昌大学 中德联合研究院,江西 南昌 330047
  • 出版日期:2013-07-25 发布日期:2013-08-02

Enrichment of Clenbuterol in Pork Using Immunomagnetic Nanobeads

LI Chao-hui,XIONG Yong-hua,GUO Liang,WU Ke-sheng,LIANG Yu-yan   

  1. 1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China;
    2. Sino-German Joint Research Insititute, Nanchang University, Nanchang 330047, China
  • Online:2013-07-25 Published:2013-08-02

摘要:

以生物素-链霉亲和素系统为介导,将克伦特罗单克隆抗体与纳米磁珠偶联制备免疫磁珠,建立特异性快速富集猪肉中盐酸克伦特罗的方法。将1mg偶联有23.9μg链霉亲和素(SA)的磁珠与220μg生物素化抗体反应,得到抗体偶联量为77.2μg/mg的免疫磁珠。免疫磁珠最佳吸附、洗脱条件为4mL 0.15mol/L盐酸在剧烈振荡条件下提取1g猪肉匀浆样品中的克伦特罗,提取液中加入0.25mg免疫磁珠在37℃,200r/min振荡吸附5~10min,100μL甲醇洗脱磁珠5min,洗脱液用PB稀释10倍后ELISA方法检测。加标实验结果表明,加标量为0.25~2ng/g时回收率大于80%,相对标准偏差为11.9%~15.7%;加标量为5ng/g时回收率降至61.2%,相对标准偏差为7.8%。

关键词: 克伦特罗, 富集, 纳米磁珠, 猪肉

Abstract:

An immunoextraction method based on anti-clenbuterol (CLE) immunomagnetic nanobeads was developed
for the rapid and specific enrichment of CLE in pork. Immunomagnetic nanobeads were prepared by coupling 77.2 μg
of biotinylated antibodies onto 1 mg of nano-magnetic beads that were coated with 23.9 μg of streptavidin. The optimal
adsorption and desorption conditions were 1 g of homogenized pork extracted with 4 mL of 0.15 mol/L HCl solution with
vigorous shaking. Then, 0.25 mg of magnetic nanobeads were added to the extract and incubated 37 ℃ for 10 min on a
shaker (200 r/min). Totally 100 μL of methanol was mixed with the immunomagnetic nanobeads for 5 min to desorb CLE.
The eluate was diluted 10 times with PBS for ELISA determination. The performance of immunomagnetic nanobeads was
evaluated by using CLE-spiked samples. The results indicated that the recovery rate of CLE at spiked levels between 0.25 ng/g
and 2 ng/g were more than 80% with relative standard deviation of 11.9% to 15.7%. However, the recovery rate of CLE was
decreased to 61.2% with relative standard deviation of 7.8% while the spiked concentration was increased to 5 ng/g.

Key words: clenbuterol, enrichment, nanomagnetic bead, pork