双模态免疫层析技术的建立及其在功能饮料中他达拉非检测中的应用

Development of dual-modal immunochromatographic assay and its application in the detection of tadalafil in functional beverages

中文关键词:  他达拉非  双模态  免疫层析  功能食品  磁性量子点

英文关键词:tadalafil  dual-mode  immunochromatography  functional food  magnetic quantum dots

中文摘要:

      目的 建立一种可同时提供视觉与荧光双信号读出的免疫层析技术; 及其在功能饮料复杂基质中, 对痕量他达拉非实现高灵敏、高准确度检测的实际应用性能。方法 本研究首先将磁性纳米颗粒与量子点相结合, 制备磁性量子点复合材料, 并将其用于标记他达拉非抗体, 构建出高灵敏度、高特异性的磁性量子点探针。同时, 通过引入预孵育步骤以有效消除基质干扰; 在实验过程中, 对抗体标记量、封闭剂种类、重悬液体系及孵育液组成等关键条件进行了系统优化。结果 实验数据显示, 该双模态免疫层析技术对他达拉非的视觉最低检出限在比色信号下达0.5 ng/mL, 在荧光信号下达1 ng/mL, 且表现出良好的特异性, 与西地那非、伐地那非等结构类似物无交叉反应现象。结论 基于磁性量子点的双模态免疫层析技术巧妙融合了磁分离技术的高净化能力与量子点的高荧光灵敏度, 具有操作简便、抗干扰性强等显著优势。该技术能够实现对功能饮品中他达拉非的现场快速筛查与初步定量, 为功能饮品非法添加物的检测提供了实用且高效的技术方案。

英文摘要:

      Objective To establish an immunochromatographic technique that can provide both visual and fluorescent signal readout at the same time. To evaluate the practical application performance of this technology for high-sensitivity and high-accuracy detection of trace tadalafil in complex matrices of energy drinks. Methods In this study, magnetic nanoparticles were first combined with quantum dots to prepare magnetic quantum dot composite materials, which were then used to label the anti-tadalafil antibody, thereby constructing a highly sensitive and specific magnetic quantum dot probe. Meanwhile, a pre-incubation step was introduced to effectively eliminate matrix interference. During the experimental process, key conditions including the antibody labeling amount, type of blocking agent, resuspension buffer system and composition of the incubation solution were systematically optimized. Results Experimental data showed that the dual-modal immunochromatographic technology achieved a visual limit of detection for tadalafil of 0.5 ng/mL in the colorimetric mode and 1 ng/mL in the fluorescent mode. Additionally, the technology exhibited excellent specificity with no cross-reactivity observed with structural analogs such as sildenafil and vardenafil. Conclusion The dual-mode immunochromatographic technique based onmagnetic quantum dots ingeniously combine the high purification capability of magnetic separation technology with the high fluorescence sensitivity of quantum dots, offering significant advantages such as simplicity of operation and strong anti-interference ability. This technique can achieve rapid screening and preliminary quantification of tadalafil in functional foods on-site, providing a practical and efficient technical solution for the detection of illegal additives in functional foods.