使用 DMAv为环境应用 DMMTAv和 DMDTAv的准备: 合成、纯化和确认
Summary
本文介绍了 dimethylmonothioarsinic 酸 (DMMTAv) 和 dimethyldithioarsinic 酸 (DMDTAv) 合成、诱导 dimethylarsinic 酸 (dmav) thiolation 通过混合 DMAv 的改进实验协议。, Na2S, 和 H2, 所以4。修改后的协议提供了实验指导, 从而克服了在定量分析过程中可能导致实验失败的综合步骤的局限性。
Abstract
Dimethylated thioarsenicals, 如 dimethylmonothioarsinic 酸 (DMMTAV) 和 dimethyldithioarsinic 酸 (DMDTAv), 是由 dimethylarsinic 酸 (DMAV) thiolation 的新陈代谢途径产生的, 最近在环境和人体器官中发现的。DMMTAv和 DMDTAv可以量化, 以确定 dimethylated thioarsenicals 的生态效应及其在环境介质中的稳定性。这些化合物的合成方法是原始社会的, 使得复制以前的研究具有挑战性。此外, 缺乏关于储存技术的信息, 包括没有物种转化的化合物的储存。此外, 由于只有有限的合成方法信息, 在合成标准化学品和进行定量分析方面可能存在实验困难。本文提出的协议为 dimethylated thioarsenicals、DMMTAv和 DMDTAv提供了一个实际改进的合成方法, 并将有助于利用高性能液体进行物种分离分析的量化。用电感耦合等离子体质谱 (HPLC-ICP) 相结合的色谱法。通过对化学试剂的制备、过滤方法和贮存的研究, 对该过程的实验步骤进行了改进。
Introduction
由于 dimethylarsinic 酸 (DMAV) 已被证明表现出急性毒性和遗传毒性由于正在接受甲基化和 thiolation 在摄取1,2, 砷 thiolation 的代谢途径有深入研究了体外和体内3,4以及环境介质 (例如,垃圾渗滤液)5,6。以前的研究在活细胞中发现了 DMAV的减少和 thiolated 类似物, 例如、 dimethylarsinous 酸 (DMAIII)、dimethylmonothioarsinic 酸 (DMMTAV) 和 dimethyldithioarsinic 酸 (DMDTAV)7,8,9, 与 dimethylated thioarsenicals 如 DMMTAV显示更大的毒性比其他已知的无机或有机砷10。高毒性 thioarsenicals 的丰度具有严重的环境影响, 因为它们在高度 sulfidic 条件下可能对人类和环境构成风险11。然而, DMMTAv和 DMDTAv (横贯) 形成的机制及其在环境媒体中的命运仍然需要进一步研究。因此, 需要对 thioarsenicals 进行定量分析, 以提高对 DMMTAv和 DMDTAv的环境影响的理解。
虽然标准化学品是定量分析的关键要求, 但由于物种转化为其他物种的风险很高, DMMTAv和 DMDTAv的标准很难通过复制以前的研究获得。原始社会合成过程12。此外, 所提到的方法有局限性, 可能会导致在合成标准化学品和进行定量分析的实际困难。DMMTAv和 DMDTAv通常是通过 dmav 2的解决方案混合 dmav、Na2和 H2, 所以4在某摩尔比率1或冒泡 H1314的气体中进行的。冒泡方法使用 H2S 气体的直接供应, 以硫磺代替氧气, 这是剧毒和难以控制的经验不足的用户。相反, 上述混合方法1, 广泛用于定性分析 DMMTAv和 DMDTAV在环境研究5,6,12, 功能 thiolation使用 H2生成的 DMAV由混合 Na2s 和 H2, 因此4和生产 DMMTAv和 DMDTAv, 允许更容易的化学计量控制生产目标化学品, 与直接使用 H2的气体。
参考混合方法过程1,3,4,8,15在本研究中提到的一些关键实验步骤的局限性, 这可能导致实验失败。例如, 具体溶剂的详细情况 (即,去离子水) 的制备和合成砷的提取和结晶是过于缩写或没有充分详细说明。有关程序性步骤的这种分散和有限的信息可能导致 thioarsenicals 的不一致形成和不可靠的量化分析。因此, 本文所开发的修改后的协议描述了 DMMTAv和 DMDTAv库存解决方案的合成, 并进行了定量的物种分离分析。
Protocol
Representative Results
Discussion
已开发的协议澄清了以前研究的关键步骤1,3,4,8,15省略或缩写, 这可能导致困难或失败期间DMMTAv和 DMDTAv合成。由于 DMMTAV是氧化敏感的1,5, 使用 N2清除的去离子水 (步骤 1.1.1-1.1.3) 来制备合成试剂, 以防…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项研究得到了基础科学研究项目 (项目编号: 2016R1A2B4013467) 的支持, 通过韩国国家研究基金会 (NRF), 由科学部、信息和通信技术 & 未来规划2016提供资金, 并得到韩国基础科学的支持。研究所研究计划 (项目编号: C36707)。
Materials
| Cacodylic acid | Sigma-Aldrich | 20835-10G-F | |
| Sodium sulfide nonahydrate | Sigma-Aldrich | S2006-500G | |
| Sulfuric acid 96% | J.T.Baker | 0000011478 | |
| Ammonium acetate | Sigma-Aldrich | A7262-500G | |
| Formic acid 98% | Wako Pure Chemical Industries, Ltd. | 066-00461 | |
| Diethyl ether (Extra Pure) | Junsei Chemical | 33475-0380 | |
| Adapter cap for 60 mL Bond Elut catridges | Agilent Technologies | 12131004 | Syringe type of SPE |
| Bond Elut C18 cartridge | Agilent Technologies | 14256031 | Syringe type of SPE |
| HyPURITY C-18 | Thermo Scientific | 22105-254630 | 5 um, 125 x 4.6 mm |
| Glovebox | Chungae-chun, Rep. of Korea | Customized | |
| Agilent 1260 Infinity Bio-inert LC | Agilent Technologies | DEAB600252, DEACH00245 | |
| Agilent Technologies 7700 Series ICP-MS | Agilent Technologies | JP12031510 | |
| Finnigan LCQ Deca XP MAX Mass Spectrometer System | Thermo Electron Corporation | LDM10627 |
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