尿样中克仑特罗免疫亲和色谱-气/质联用检测方法的研究

Determination of Clenbuterol in Urine Using Immunoaffinity Chromatographic Cleanup and Gas Chromatography-Mass Spectromety

以单抗为配基制备克仑特罗免疫亲和柱,建立了简单、快速、特异的样品净化技术,柱容量达到100 ng。偶联抗体量与柱容量呈线性关系,线性方程为:y=236.36x+6.6214,R2=0.9461。测定尿样中的克仑特罗,不需前处理直接用亲和柱提纯后以气质联用(GC/MS)方法检测。空白猪尿添加浓度为205、01、00 ng时,回收率分别为85%±1.3,85.9%±1.3,81.5%±2.7。同时以NY/XQ421-2003标准中净化方法为对照,回收率分别为113%±5.6,94.5%±10,78.6%±1.7,偶联非特异抗体的对照组亲和柱对样品保留值为7.03%。数据显示:免疫亲和柱作为提纯方法,与传统固相萃取提纯方法相比平行性好,提纯后样本杂质少。因此免疫亲和柱是净化样品的一个有效方法。

A simple, rapid and specific preparative method was developed, which was based on monoclonal antibody-mediated immunoaffinity column （IAC） for clenbuterol. The dynamic column capacity was more than 100 ng , which is related with the antibody quantity, and the regression equation was y= 236. 36x ＋ 6. 6214,R^2= 0. 9461.Urine sample could be directely applied to immunoaffinity column before determination by GC/MS without any other pretreatments. The recoveries of blank pig urine fortified samples at levels of 20 ng/mL,50 ng/mL and 100 ng/mL were 85%±1.3,85.9%±1.3 and 81.5%±2.7. At the same time, to treat the fortified urine samples by the standard method of NY/XQ421-2003, the recoveries were 113%±5.6,94.5%±10 and 78.6%±1.7. A control column prepared from nonspecific IgG retained 7. 03%. The results showed that compared to traditional solid phase extraction （SPE） cleanup method, IAC has a better parallelism and reduces more impurities which facilitates immunoaffinity chromatography to be an efficient cleanup method.