氯霉素免疫亲和柱的制备及在牛奶中痕量氯霉素残留分析中的应用  

PREPARATION OF IMMUNOAFFINITY COLUMN OF CHLORAMPHENICOLFOR TRACE ANALYSISOF CHLORAMPHENICOL IN MILK

作　　者：乔好[1] 赵立杰[1] 张岩[1] 徐蓓[1] 王凤侠[1] 王硕[1] 

机构地区：[1]天津科技大学食品营养与安全省部共建教育部重点实验室,天津300457

出　　处：《食品研究与开发》2008年第4期141-145,共5页Food Research and Development

基　　金：科技部"十一五"国家科技支撑计划(2006BAD05A06)

摘　　要：将氯霉素多克隆抗体与sepharose-4B偶联,制备了对氯霉素有特异性吸附的免疫亲和柱。采用紫外扫描鉴定偶联反应,利用直接竞争酶联免疫吸附法及高效液相色谱法对制备好的免疫亲合柱进行评价。通过纯化牛奶基质,对该免疫亲和柱的纯化能力进行了研究。结果表明,12.84mg抗体与sepharose-4B的联接率为91.06%。以2%甲醇/水溶液预洗涤杂质,80%甲醇/水溶液洗脱分析物,可以消除牛奶基质对高效液相色谱分析氯霉素的影响。在牛奶中添加1.5μg/L～5μg/L氯霉素标品,测得其回收率为95.2%～105.9%,相对标准偏差为1.19%～4.18%。该亲和柱可以明显消除基质影响,提高检测的灵敏度。A immuoaffinity column was prepared by coupling anti-chloramphenicol muhiclonal antibody with sepharose-4B. After determining the coupling rate by UV absorbance measurement, the coulumn has been evaluated through direct-competetion ELISA and HPLC. Then the purification capacity of the IAC has been tested by using milk as a sample. 12.84 mg antibody was coupled with sepharose-4B which produced a coupling rate of 91.06 %. Use of 2 % methanol/water buffer as washing buffer and 80 % methanol/water as elution buffer can eliminate the impact milk brings to the test by HPLC. The mean recoveries were in the range 95.2 %- 105.9 % and RSD was 1.19 %-4.18% at levels of 1.5 μg/L- 5 μg/L.

关 键 词：免疫亲和柱 氯霉素 牛奶 高效液相 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]