纳米免疫磁珠富集猪肉中的盐酸克伦特罗  

Enrichment of Clenbuterol in Pork Using Immunomagnetic Nanobeads

作　　者：李超辉[1,2] 熊勇华[1,2] 郭亮[1,2] 吴科盛[2] 梁语嫣[2] 

机构地区：[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　　处：《食品科学》2013年第14期182-186,共5页Food Science

基　　金："十二五"国家科技支撑计划项目(2011BAK10B06;2011BAK10B04);江西省科技厅自然科学基金项目(2010GZN0143);南昌大学分析测试基金项目(2012028)

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%。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.

关 键 词：克伦特罗 富集 纳米磁珠 猪肉 

分 类 号：TS201.6[轻工技术与工程—食品科学]