Li Yingguo, Chen Yiqiang, Li Zhengguo, Zhang Lei, Li Xianliang, Xi Cunxian, Wang Guomin, Wang Xiong, Guo Qi, Li Na
College of Bioengineering, Chongqing University, Chongqing, China.
J Chromatogr Sci. 2012 Mar;50(3):167-74. doi: 10.1093/chromsci/bms006.
This paper describes the preparation of a novel mixed-bed immunoaffinity chromatography (IAC) column by coupling four monoclonal antibodies against different sulfonamides (SAs) to Sepharose 4B. The IAC column can be used to simultaneously extract and purify 16 SAs in pork muscle. The dynamic column capacities for all SAs in mixed standard solution were between 312 and 479 ng/mL gel. After simple extraction and IAC cleanup, the sample solution can be directly injected for liquid chromatography-ultraviolet analysis. The recoveries of SAs from spiked samples at levels of 25, 50 and 100 µg/kg ranged from 83.3 to 103.1% with variation coefficient less than 8.6%. The comparison of IAC with liquid-liquid extraction and solid phase extraction indicated that IAC has better purification effect and needs less organic solution than conventional methods, thus it would be an ideal method for selective purification of SAs in pork muscle.
本文描述了一种新型混合床免疫亲和色谱(IAC)柱的制备方法,该方法是将四种针对不同磺胺类药物(SAs)的单克隆抗体偶联到琼脂糖4B上。该IAC柱可用于同时提取和纯化猪肉肌肉中的16种磺胺类药物。混合标准溶液中所有磺胺类药物的动态柱容量在312至479 ng/mL凝胶之间。经过简单提取和IAC净化后,样品溶液可直接进样进行液相色谱-紫外分析。在25、50和100 µg/kg加标水平下,磺胺类药物的回收率在83.3%至103.1%之间,变异系数小于8.6%。IAC与液液萃取和固相萃取的比较表明,IAC具有更好的净化效果,且比传统方法需要更少的有机溶液,因此它将是一种选择性纯化猪肉肌肉中磺胺类药物的理想方法。