Zhang Can, Wang Shuo, Fang Guozhen, Zhang Yan, Jiang Liqin
Key Laboratory of Food Nutrition and Safety, Ministry of Education, Tianjin University of Science and Technology, Tianjin, P.R. China.
Electrophoresis. 2008 Aug;29(16):3422-8. doi: 10.1002/elps.200800188.
A competitive immunoassay using CE with an LIF detector was developed for the detection of chloramphenicol (CAP). The method was based on the competitive reactions between fluorescently labeled CAP hapten and free CAP, with a limited amount of anti-CAP antibody. The poly(N-isopropylacrylamide) (pNIPA) hydrogel was added in the separation buffer as a dynamic modifier to reduce adsorption and enhance reproducibility. The linear range and LOD for CAP were 0.008-5 microg/L and 0.0016 microg/L, respectively. An ELISA using the same immuno-reagents was also developed for the analysis of CAP, with an LOD of 0.03 microg/L. The sensitivity of this CE immunoassay (CEIA)-LIF was almost 20 times greater than that of the ELISA. Using CEIA-LIF, equilibrium was reached in 15 min and the analytical results were obtained within 5 min by CE separation. Sample preparation for CEIA-LIF was not time-consuming and the matrix effect was easy to remove. An LOD of 0.1 microg/kg CAP in food matrices was easily achieved. This method is thus proposed as a fast and sensitive means of detecting trace amounts of CAP residues in animal-derived foods.
开发了一种使用毛细管电泳（CE）结合激光诱导荧光（LIF）检测器的竞争性免疫分析法用于检测氯霉素（CAP）。该方法基于荧光标记的CAP半抗原与游离CAP之间的竞争性反应，以及有限量的抗CAP抗体。在分离缓冲液中添加聚（N-异丙基丙烯酰胺）（pNIPA）水凝胶作为动态修饰剂，以减少吸附并提高重现性。CAP的线性范围和检测限分别为0.008 - 5μg/L和0.0016μg/L。还开发了一种使用相同免疫试剂的酶联免疫吸附测定（ELISA）用于CAP分析，其检测限为0.03μg/L。这种毛细管电泳免疫分析法（CEIA）-LIF的灵敏度几乎是ELISA的20倍。使用CEIA-LIF，15分钟内达到平衡，通过CE分离在5分钟内获得分析结果。CEIA-LIF的样品制备不耗时，且基质效应易于消除。在食品基质中轻松实现了0.1μg/kg CAP的检测限。因此，该方法被提议作为一种快速、灵敏的检测动物源性食品中痕量CAP残留的手段。
