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首页> 外文期刊>Journal of mass spectrometry: JMS >Rapid and sensitive identification of ricin in environmental samples based on lactamyl agarose beads using LC-MS/MS (MRM)
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Rapid and sensitive identification of ricin in environmental samples based on lactamyl agarose beads using LC-MS/MS (MRM)

机译：使用LC-MS / MS（MRM）基于内酰亚胺琼脂糖珠的环境样品中蓖麻植物的快速和敏感鉴定

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摘要

Ricin, a plant-derived toxin extracted from the seeds of Ricinus communis (castor bean plant), is one of the most toxic proteins known. Ricin's high toxicity, widespread availability, and ease of its extraction make it a potential agent for bioterrorist attacks. Most ricin detection methods are based on immunoassays. These methods may suffer from low efficiency in matrices containing interfering substances, or from false positive results due to antibody cross reactivity, with highly homologous proteins. In this study, we have developed a simple, rapid, sensitive, and selective mass spectrometry assay, for the identification of ricin in complex environmental samples. This assay involves three main stages: (a) Ricin affinity capture by commercial lactamyl-agarose (LA) beads. (b) Tryptic digestion. (c) LC-MS/MS (MRM) analysis of tryptic fragments. The assay was validated using 60 diverse environmental samples such as soil, asphalt, and vegetation, taken from various geographic regions. The assay's selectivity was established in the presence of high concentrations of competing lectin interferences. Based on our findings, we have defined strict criteria for unambiguous identification of ricin. Our novel method, which combines affinity capture beads followed by MRM-based analysis, enabled the identification of 1 ppb ricin spiked into complex environmental matrices. This methodology has the potential to be extended for the identification of ricin in body fluids from individuals exposed (deliberately or accidentally) to the toxin, contaminated food or for the detection of the entire family of RIP-II toxins, by applying multiplex format.
机译：来自Ricinus Communis（蓖麻植物）的种子中提取的植物衍生的毒素是最有毒的蛋白质之一。丽林的毒性高，可用性广泛，其提取易于提取使其成为生物恐怖主义攻击的潜在代理。大多数Ricin检测方法基于免疫测定。这些方法可能在含有干扰物质的基质中患有低效率，或由于抗体交叉反应性引起的假阳性结果，具有高度同源的蛋白质。在这项研究中，我们开发了一种简单，快速，敏感和选择性的质谱法，用于鉴定蓖麻植物中的蓖麻植物。该测定涉及三个主要阶段：（a）通过商业内人酰基 - 琼脂糖（La）珠子捕获蓖麻植物。 （b）胰蛋白酶消化。 （c）LC-MS / MS（MRM）对胰蛋白酶片段的分析。使用60种不同的环境样品（如土壤，沥青和植被）验证了测定，从各种地理区域取出。在存在高浓度的竞争凝集素干扰的情况下建立了测定的选择性。根据我们的研究结果，我们确定了明确识别Ricin的严格标准。我们的新方法结合了亲和捕获珠，其次是基于MRM的分析，使鉴定为1ppb蓖麻毒素掺入复杂的环境基质中。该方法具有延伸潜力，用于通过施加多路复用格式，从暴露（故意或意外地）暴露（故意或意外）的个体的体液中的蓖麻液中的蓖麻液中的蓖麻液中的刺激。

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