Instituto Universitario de Reconocimiento Molecular y Desarrollo Tecnológico, Departamento de Química, Universidad Politécnica de Valencia, camino de vera s/n E46022, Valencia, Spain.
Environ Sci Technol. 2010 Dec 1;44(23):9024-9. doi: 10.1021/es101653r. Epub 2010 Nov 3.
The development of simple, accurate, and rapid multisample analytical methodologies to find out critical targets in waters is highly demanded. Optical microsensor arrays to determine microcystins in river waters are developed on the polycarbonate side of compact discs. The working principle of the sensors relied on an indirect competitive microimmunoassay, where free microcystin LR (MC-LR) competes with immobilized conjugate for specific monoclonal antibody. The results of the immunoreaction are detected with a DVD drive, showing the readouts in minutes. The method reached a sensitivity (IC(50)) for MC-LR of 1.04 μg/L and a linear response in the range 0.12-2.00 μg/L, allowing its determination below the upper limit proposed by the World Health Organization in drinking water. The developed analytical approach shows simplicity, good sensitivity, high throughput capability, and rapidity (37 min) in field use. The optimized assay showed also high congener reactivity to MC-LY (144%), MC-LA (125%), MC-LF (119%), MC-LW (102%), MC-YR (83%), and nodularin (94%). Furthermore, the suitability of the disk biosensor to quantify MC-LR was successfully evaluated analyzing river water samples, obtaining excellent recoveries (78-113%). Precoated discs are stable for at least seven weeks without loosing their analytical performances. Also, the portability of the analytical system permits on-site analysis and quantification, saving time and other resources. To our knowledge, this is the only work where a portable, easy-to-use, array based system has been developed for on-site microcystin quantification and applied to simultaneously analyze 42 samples plus the calibration curve, reaching microgram per liter sensitivity.
开发简单、准确、快速的多水样分析方法来检测水中的关键目标是非常有必要的。在光盘的聚碳酸酯一侧开发了用于测定河水中微囊藻毒素的光学微传感器阵列。传感器的工作原理依赖于间接竞争微免疫测定法，其中游离微囊藻毒素 LR（MC-LR）与固定化缀合物竞争与特异性单克隆抗体结合。免疫反应的结果用 DVD 驱动器检测，几分钟内即可得到读数。该方法对 MC-LR 的检测灵敏度（IC50）为 1.04μg/L，线性响应范围为 0.12-2.00μg/L，允许其在世界卫生组织规定的饮用水上限以下进行测定。所开发的分析方法具有简单、高灵敏度、高通量能力和快速（37 分钟）的特点，适用于现场使用。优化后的测定法对 MC-LY（144%）、MC-LA（125%）、MC-LF（119%）、MC-LW（102%）、MC-YR（83%）和节球藻毒素（94%）也具有较高的同系物反应性。此外，圆盘生物传感器定量 MC-LR 的适用性通过分析河水样本得到了成功评估，获得了极好的回收率（78-113%）。预涂覆的光盘至少稳定七周而不会失去其分析性能。此外，该分析系统的便携性允许现场分析和定量，节省了时间和其他资源。据我们所知，这是唯一一项开发了便携式、易于使用的基于阵列系统用于现场微囊藻毒素定量，并应用于同时分析 42 个样本加校准曲线，达到微克/升灵敏度的工作。
