Novel Natural and Synthetic Brominated and Iodinated Compounds in the Environment

该研究旨在通过使用效应导向分馏和鉴定来确定新型化合物的生物活性，从而继续鉴定新型有机溴（OBC）和有机碘（OIC）化合物。 包括活细胞阵列在内的一系列功能性生物测定将用于鉴定受最活跃的化合物类别影响的分子途径。 将联合收割机超高分辨率质谱与蛋白质组学相结合的“下拉”方法将被开发用于鉴定新型OBCs和OI的分子靶点。 拟议的研究有几个具体目标：1）识别和量化一系列海洋和淡水环境中的新型OBCs和OIC; 2）使用质量平衡（溴和碘）方法来确定可能存在哪些额外的OBCs和OIC，以及3）使用生物测定来确定新型OBCs和OIC的毒性和潜在有益用途。 拟议的研究计划将首先集中在食物链的基础上，包括海藻和海绵和珊瑚等无脊椎动物。OBC将通过靶向和非靶向质谱法的组合进行鉴定，包括超高分辨率质谱法UHR LC/MS（OrbiTrap）和三重四极杆串联质谱法（LC-MS/MS）、LC四极杆-飞行时间质谱法（Q-ToF）和高分辨率GC高分辨率质谱法（HRGC-HRMS）。 将通过仪器中子活化分析（INAA）测定粗样品或有机溶剂浸提液的总溴（TBr）和总碘（TI）。 将通过M1确定准确质量和分子式，并使用M2数据阐明结构。将确定已知的合成OBC和OIC以及建议作为替代阻燃剂的化合物的浓度。 将提取足量的未知化合物，并通过制备型HPLC进行纯化。 然后将浓缩的分离物进行标准方法，如UV和光谱法和IR光谱法和元素分析，以帮助阐明和/或确认结构。 一旦结构得到确认，就可以开发合成程序并制备更大量的纯化合物。 一旦获得更大质量的材料，将对其进行一系列生物测定以确定：1）其毒性作用模式和效应阈值，以及第二组生物测定以确定其作为治疗剂的潜在效用。 除了筛选毒性和潜在的有益用途外，还将调查其生产的潜在生态和/或生理原因。 这将通过使用微生物培养物的逆向工程筛选来完成，基于特定化合物合成途径中酶的存在或不存在。 如果化合物被确定为毒性太大，则不会进一步测试它们的治疗特性。

The research is designed to continue to identify novel organo-bromine (OBCs) and organo-iodine (OICs) compounds by use of effect-directed fractionation and identification to determine biological activities of novel compounds. A range of functional bioassays including live cell arrays will be used to identify molecular pathways affected by the most active classes of compounds. A “pull-down” approach that will combine ultra-high resolution mass spectrometry with proteomics will be developed to identify molecular targets of novel OBCs and OIs. The proposed research has several specific objectives: 1) identify and quantify novel OBCs and OICs in a range of marine and freshwater environments; 2) use a mass balance (on Br and I) approach to determine what additional OBCs and OICs might be present and 3) use bioassays to determine the toxicity and potential beneficial uses of novel OBCs and OICs. The proposed research program will focus initially on the base of the food chain, including marine algae and invertebrates such as sponges and corals. OBCs will be identified by use of a combination of targeted and un-targeted mass spectrometry, including ultra-high resolution mass spectrometry UHR LC/MS (OrbiTrap) and triple quadrupole tandem mass spectrometry (LC-MS/MS), LC quadrupole-time of flight mass spectrometry (Q-ToF) and high resolution GC High Resolution Mass spectrometry (HRGC-HRMS). Total Br (TBr) and total iodine (TI) will be determined by Instrumental Neutron Activation Analysis (INAA) of either crude samples or organic solvent extracts. Accurate masses and formulas will be determined by M1 and structures elucidated by use of M2 data. Concentrations of known synthetic OBCs and OICs and those compounds that have been suggested as replacement flame retardants will be determined. Sufficient quantities of unknown compounds will be extracted and purified by preparative HPLC. The concentrated isolates will then be subjected to standard methods such as UV and spectrometry and IR spectroscopy and elemental analyses to help elucidate and or confirm structures. Once a structure has been confirmed, a synthesis procedure can be developed and greater quantities of pure compounds made. Once greater masses of material are available they will be subjected to a range of bioassays to determine: 1) their toxic mode of action and threshold for effects and a second set of bioassays to determine their potential utility as therapeutic agents. In addition to screening for toxicity and potential beneficial uses, the potential ecological and or physiological reasons for their production will be investigated. This will be done by use of a reverse-engineering screening of microbial cultures, based on the presence or absence of enzymes in pathways of syntheses of specific compounds. If compounds are determined to be too toxic, they will not be tested farther for therapeutic properties.