Synthesis and quantitation of microcystins and anabaenopeptins in the Great Lakes region to establish human exposure risks via ingestion and inhalation

项目摘要 气候变化、农业实践、水文变化和污水排放都导致了 在蓝藻有害藻华（cHABs）的频率和强度，产生生物活性和毒性， 次级肽代谢物。因此，公共卫生和饮用水供应，娱乐用水 关闭，沿海海产品污染增加。蓝细菌肽毒素是已知的原因 或疑似肝脏、神经系统、皮肤、胃肠道和肾脏疾病。尽管数百个潜在的 有毒和有益的生物活性肽已经报道，只有少数参考材料， 市售。cHAB肽的研究、监测和缓解一直受到缺乏 认证和散装参考物质。分析纯（经认证）合成标准品氰毒素和 为保护公众健康，迫切需要可靠的分析工作流程。我们的长期目标是创造 稳定可靠的CHAB氰肽参比物质供应，可用于氰肽的研究。 发生和人类健康影响。我们的目标是建立有效的合成路线 鱼腥藻肽（ABP）和微囊藻毒素（MCs）的生物活性谱，以创建质谱 工作流程，以确定其识别和量化，并确定摄入和吸入的影响 作为暴露途径。特别是，我们将评估蓝藻毒素对呼吸道细胞的影响 和重建的胃肠道微生物组。在五大湖地区的淡水中观察到ABP和MC 高浓度，已知有数百种具有不同毒性程度的同系物 和生物活性。这些湖泊/河流中有许多含有暂时或未指定的同源物。我们 同源物选择旨在提高靶向和非靶向质量的可靠性和再现性 光谱分析工作流程。了解蓝藻毒素的暴露途径和健康风险取决于 化学结构及其与环境的相互作用。我们提出的气溶胶研究将有助于确定 与吸入有关的风险。迫切需要合成氰基肽/同系物， 进行了表征和系统研究，以量化其在cHAB水中的相关暴露风险， 气溶胶我们提出的工作结合了化学和药理学几个领域的专业知识， 重要的是，它的进步使正在进行的基础cHAB研究在生态学领域， 毒理学，生物学，命运和运输，并通过认证提供可靠的氰毒素定量 水资源管理者和公共卫生决策者的标准。

PROJECT ABSTRACT Climate change, agriculture practices, hydrology alterations, and sewage sheds are contributing to an increase in the frequency and intensity of cyanobacteria harmful algal blooms (cHABs) that produce bioactive and toxic secondary peptide metabolites. As a result, public health and drinking water advisories, recreational water closures, and coastal seafood contamination have increased. Cyanobacteria peptide toxins are known causes or suspect in liver, neurological, dermal, gastrointestinal and kidney disease. Although hundreds of potential toxic and beneficial bioactive peptides have been reported, only a handful of reference materials are commercially available. cHAB peptide research, monitoring, and mitigation has been bottlenecked by lack of certified and bulk reference materials. Analytically pure (certified) synthetic standards cyanotoxin and reliable analytical workflows are critically needed to protect public health. Our long-term goal is to create a stable and reliable supply of CHAB cyanopeptide reference materials to enable investigation of cyanopeptides’ occurrence and human health impacts. Our objectives in this proposal are to establish efficient synthetic routes and bioactivity profiles for anabaenopeptins (ABPs) and microcystins (MCs), to create mass spectrometry workflows for their identification and quantification, and to establish the impact of ingestion and inhalation pathways as exposure routes. In particular, we will assess cyanotoxins’ impact on cells from the respiratory tract and on reconstituted GI microbiomes. The ABPs and MCs are observed in the Great Lakes region’s freshwater bodies in high concentrations and are known to have hundred(s) of congeners with varying degrees of toxicity and bioactivity. Many of these lakes/rivers contain congeners with tentative or unassigned identifications. Our congener selection is designed to increase the reliability and reproducibility of targeted and untargeted mass spectrometry workflows. Understanding the cyanotoxin exposure routes and health risk is dependent on chemical structure and its interplay with the environment. Our proposed aerosol studies will assist in determining the risk associated with inhalation. There is a critical need for cyanopeptides/congeners to be synthesized, characterized, and systematically studied to quantify their associated exposure risks in cHAB water and aerosols. Our proposed work combines the expertise of several areas of chemistry and pharmacology and is important because it advances empowers ongoing fundamental cHAB research in the areas of ecology, toxicology, biology, and fate and transport, and provides reliable cyanotoxin quantification through certified standards for water resource managers and public health decision makers.