Yan Long, Li Kexuan, Wu Jina, Xing Zhongfang, Li Xiaosen, Liu Shilei
State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China.
Int J Mol Sci. 2025 Jun 12;26(12):5641. doi: 10.3390/ijms26125641.
As a type II ribosome-inactivating protein (RIP-II) toxin, Ricin has garnered widespread recognition due to its inherent qualities as an easily prepared and highly stable substance, posing serious implications as a potential chemical and biological terrorist threat. For the detection of ricin, traditional immunoassay technologies, including methods like peptide cleavage combined with liquid chromatography mass spectrometry (LC-MS) or the more commonly used enzyme-linked immunosorbent assay (ELISA), have offered reliable results. However, these techniques are unfortunately limited by the requirement of a complex sample pretreatment process, which can be time-consuming and labor-intensive. In an effort to overcome these limitations, a highly sensitive and selective method was introduced via metal element labeling combined with inductively coupled plasma mass spectrometry (ICP-MS) in this research. The method centered on designing and synthesizing a europium-labeled compound (DOTA-NHS-Eu) that specifically targets the amino groups (-NH) on ricin. The compound, coupled with the application of specific magnetic beads, achieved the specific enrichment and subsequent quantitative detection of ricin by ICP-MS, which is based on the amount of europium element present. The established method demonstrated high specificity for ricin recognition, with a signal response to bovine serum protein that was found to be less than 10% of that for ricin. Furthermore, the calibration curve created for the method (y = 81.543x + 674.02 (R > 0.99)) for quantifying ricin in a concentration range of 1.0-100 μg/mL demonstrated good linearity. The method was further evidenced by the limit of detection and quantitation results of 0.1 and 1.89 μg/mL, respectively. Collectively, these findings suggested that the research has offered a highly sensitive and selective method for ricin detection, which was not only easy to operate but also provided efficient results. The scheme showed great potential for the verification of chemical weapons and the destruction of toxic chemicals, therefore representing a significant advancement in the field of biomolecular detection and analysis.
作为一种II型核糖体失活蛋白（RIP-II）毒素，蓖麻毒素因其易于制备且高度稳定的内在特性而广受关注，这使其成为潜在的化学和生物恐怖主义威胁，具有严重影响。对于蓖麻毒素的检测，传统免疫分析技术，包括肽段裂解结合液相色谱质谱联用（LC-MS）等方法或更常用的酶联免疫吸附测定（ELISA），都能提供可靠结果。然而，遗憾的是，这些技术受到复杂样品预处理过程的限制，该过程既耗时又费力。为了克服这些限制，本研究通过金属元素标记结合电感耦合等离子体质谱（ICP-MS）引入了一种高灵敏度和选择性的方法。该方法的核心是设计并合成一种铕标记化合物（DOTA-NHS-Eu），它能特异性靶向蓖麻毒素上的氨基（-NH）。该化合物与特定磁珠的应用相结合，通过基于铕元素含量的ICP-MS实现了蓖麻毒素的特异性富集和后续定量检测。所建立的方法对蓖麻毒素识别具有高特异性，对牛血清蛋白的信号响应不到蓖麻毒素的10%。此外，该方法在1.0 - 100 μg/mL浓度范围内定量蓖麻毒素的校准曲线（y = 81.543x + 674.02（R > 0.99））显示出良好的线性。该方法的检测限和定量限结果分别为0.1和1.89 μg/mL，进一步证明了该方法的有效性。总体而言，这些发现表明该研究为蓖麻毒素检测提供了一种高灵敏度和选择性的方法，不仅操作简便，而且结果高效。该方案在化学武器核查和有毒化学品销毁方面显示出巨大潜力，因此代表了生物分子检测与分析领域的一项重大进展。
