van de Water C, Tebbal D, Haagsma N
Faculty of Veterinary Medicine, Department of the Science of Food of Animal Origin, University of Utrecht, The Netherlands.
J Chromatogr. 1989 Sep 8;478(1):205-15. doi: 10.1016/s0021-9673(01)84386-7.
A simple, rapid and specific sample preparation method based on antibody-mediated clean-up for the determination of chloramphenicol (CAP) in milk and eggs was developed. Skimmed milk and centrifuged egg homogenates were filtered and directly applied to immunoaffinity columns which were prepared by coupling monoclonal antibodies against CAP to a carbonyldiimidazole-activated support. Using a 0.2 M glycine, 0.5 M NaCl (pH 2.8) solution as an eluent, the immunoaffinity columns can be used more than 30 times without a decrease in column capacity. In subsequent high-performance liquid chromatographic analysis, no matrix interferences were observed. Good recoveries were obtained at spiking levels of 1-100 micrograms kg-1. Due to the high specificity of the clean-up procedure, the limit of detection can be lowered by increasing the test portion. Concerning milk, the limit of detection was successfully lowered to 20 ng kg-1 by increasing the test portion to 11 (recovery 99%). The method was applied to eggs produced by hens treated with CAP. The results are compared with those obtained by solid-phase extraction using silica gel.
建立了一种基于抗体介导净化的简单、快速且特异的样品制备方法，用于测定牛奶和鸡蛋中的氯霉素（CAP）。脱脂牛奶和离心后的鸡蛋匀浆经过过滤后直接应用于免疫亲和柱，该免疫亲和柱是通过将抗CAP单克隆抗体偶联到羰基二咪唑活化的载体上制备而成。使用0.2 M甘氨酸、0.5 M氯化钠（pH 2.8）溶液作为洗脱剂，该免疫亲和柱可重复使用30多次且柱容量无下降。在随后的高效液相色谱分析中，未观察到基质干扰。在1 - 100微克/千克的加标水平下获得了良好的回收率。由于净化程序的高特异性，可通过增加测试部分来降低检测限。对于牛奶，通过将测试部分增加到11（回收率99%），成功将检测限降低到了20纳克/千克。该方法应用于用CAP处理过的母鸡所产鸡蛋的检测。并将结果与使用硅胶进行固相萃取所获得的结果进行了比较。
