Novel Natural and Synthetic Brominated and Iodinated Compounds in the Environment

本研究旨在通过效应导向分离和鉴定来继续鉴定新型有机溴(OBCs)和有机碘(OICs)化合物，以确定新化合物的生物活性。包括活细胞阵列在内的一系列功能生物分析将被用来识别受最活跃的化合物类别影响的分子途径。将开发一种将超高分辨率质谱学与蛋白质组学相结合的“下拉”方法，以识别新的OBC和OIS的分子靶标。拟议的研究有几个具体目标：1)在一系列海洋和淡水环境中识别和量化新的OBC和OIC；2)使用质量平衡方法(在BR和I上)确定可能存在哪些额外的OBC和OIC；3)使用生物测定来确定新的OBC和OIC的毒性和潜在的有益用途。拟议的研究计划首先将重点放在食物链的基础上，包括海藻和海绵和珊瑚等无脊椎动物。将结合使用靶向和非靶向质谱学，包括超高分辨率质谱学UHR LC/MS(Orbitrap)和三重四极杆串联质谱仪(LC-MS/MS)、LC四极杆飞行时间质谱仪(Q-ToF)和高分辨率GC高分辨率质谱仪(HRGC-HRMS)。总溴(TBR)和总碘(TI)将通过仪器中子活化分析(INAA)对粗品或有机溶剂萃取物进行测定。准确的质量和公式将由M1确定，结构由M2数据阐明。已知的合成有机化合物和有机化合物以及那些被建议作为替代阻燃剂的化合物的浓度将被确定。通过制备型高效液相色谱分离纯化出大量的未知化合物。然后对浓缩的分离株进行标准方法，如紫外可见光谱、红外光谱和元素分析，以帮助阐明和或确认结构。一旦确定了结构，就可以开发合成程序并制造更多的纯化合物。一旦有更多的材料可用，它们将接受一系列生物测试，以确定：1)它们的毒性作用模式和影响阈值，以及第二套生物测试，以确定它们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，还将调查它们产生的潜在生态和/或生理原因。这将通过使用微生物培养的反向工程筛选来完成，该筛选基于特定化合物的合成途径中是否存在酶。如果化合物被确定为毒性太大，它们将不会进行进一步的治疗特性测试。

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.