An electrochemically based antibody immobilization was used to perform environmentally and clinically relevant immunoassays for stress hormones biomarkers (cortisol and cortisone) using chemiluminescence (CL) detection. To achieve CL detection, the ferrocene tag on the antibodies was first oxidised, and this then acted as a catalyst for the luminol and hydrogen peroxide CL reaction. The conditions were optimised and measurements were made with an incubation time of 30 min. Using this approach limits of detection were obtained of 0.47 pg ml−1 and 0.34 pg ml−1 also R2 0.9912 and 0.9902 for cortisol and cortisone respectively with a linear concentration from 0 to 50 ng ml−1. The method was then applied to Zebrafish whole body and artificial saliva samples. For the Zebrafish sample recoveries of 91.0% and 90.0% were obtained with samples spiked with cortisol and cortisone, for artificial saliva the recoveries were 92.59% and 90.73% respectively. Interference studies showed only minor effects on the measurement of the analyte. A comparison between this procedure and the standard enzyme-linked immunosorbent assay (ELISA) procedure gave approximately the same R2 values.

中文翻译：

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一种灵敏的基于化学发光的免疫测定法，用于检测作为应激生物标志物的皮质醇和可的松

基于电化学的抗体固定化用于使用化学发光 (CL) 检测对应激激素生物标志物（皮质醇和可的松）进行环境和临床相关的免疫测定。为了实现 CL 检测，抗体上的二茂铁标签首先被氧化，然后作为鲁米诺和过氧化氢 CL 反应的催化剂。优化条件并在 30 分钟的孵育时间下进行测量。使用这种方法获得的检测限为 0.47 pg ml-1 和 0.34 pg ml-1，皮质醇和可的松的 R2 分别为 0.9912 和 0.9902，线性浓度为 0 到 50 ng ml-1。然后将该方法应用于斑马鱼全身和人工唾液样本。对于 91.0% 和 90% 的斑马鱼样品回收率。0% 的样品加标了皮质醇和可的松，人工唾液的回收率分别为 92.59% 和 90.73%。干扰研究表明对分析物的测量只有很小的影响。该程序与标准酶联免疫吸附测定 (ELISA) 程序之间的比较给出了大致相同的 R2 值。