School of Science, Qingdao Technological University, Qingdao 266033, China.
Toxicon. 2012 May;59(6):626-32. doi: 10.1016/j.toxicon.2012.02.011. Epub 2012 Mar 1.
A new method was developed to quantify domoic acid by capillary electrophoresis-based enzyme immunoassay (CE-EIA) with electrochemical (EC) detection. The method was based on noncompetitive immunoreaction between free domoic acid antigen (Ag) and excessive amount of horseradish peroxidase (HRP)-labeled antidomoic acid antibody tracer (Ab*) in liquid phase. Then the bound enzyme-labeled complex (Ab*-Ag) and unbound Ab* were separated by CE, and the system of HRP catalyzing H(2)O(2)/o-aminophenol (OAP) reaction was adopted for EC detection. Using CE-EIA with EC detection, equilibrium was reached in 30 min, and the analytical results were obtained within a further 5 min. The linear range and the detection limit were 0.1-50 ng/mL and 0.02 ng/mL, respectively. Analytes recoveries were 89.6-105.8%. The sensitivity of the method was 16 times greater than that of enzyme-linked immunosorbent assay (ELISA). The developed method was applied to quantitatively analysis of domoic acid in contaminated shellfish samples with rapid and simple pretreatment, and the results were consistent with the same samples analyzed through ELISA. The CE-EIA with EC detection provides a valid and sensitive analytical approach, not previously available, for determination of domoic acid in shellfish samples.
建立了一种基于毛细管电泳-酶免疫分析（CE-EIA）结合电化学（EC）检测方法用于定量检测贝类中的软骨藻酸。该方法基于游离软骨藻酸抗原（Ag）与过量辣根过氧化物酶（HRP）标记的抗软骨藻酸抗体示踪剂（Ab*）在液相中的非竞争免疫反应。然后通过 CE 分离结合的酶标记复合物（Ab*-Ag）和未结合的 Ab*，并采用 HRP 催化 H（2）O（2）/邻氨基酚（OAP）反应的体系进行 EC 检测。采用 CE-EIA 结合 EC 检测，平衡在 30 min 内达到，进一步 5 min 内即可获得分析结果。线性范围和检测限分别为 0.1-50 ng/mL 和 0.02 ng/mL。分析物回收率为 89.6-105.8%。该方法的灵敏度比酶联免疫吸附法（ELISA）高 16 倍。建立的方法用于贝类样品中软骨藻酸的快速简单预处理定量分析，与 ELISA 分析相同样品的结果一致。CE-EIA 结合 EC 检测为贝类样品中软骨藻酸的检测提供了一种以前不可用的有效和灵敏的分析方法。
