Development of an interface for the on-line coupling of HPLC and HR-cs-GFAAS for fluorine speciation analysis

在药物领域，有机分子的氟化导致了新的、有前途的性质（例如改变药物的代谢）。氟化物质在技术应用领域也是有益的。不断增长的产量和应用领域导致（水生）环境中这些物质的增加。由于它们的新特性，最有可能在环境中具有不同的分布、物种形成以及生物利用度。包括高灵敏度、氟选择性和氟非特异性检测器响应的分析技术尚不具备。因此，该项目的主要目标是发展一种补充方法，使氟形态分析能够用于各种应用，包括分析（水生）环境样品，以评估含氟化合物的负荷情况、分布和生物利用度以及形态。从这种方法中获益的其他领域包括制药、医学和材料科学。高分辨率-连续源-石墨炉原子吸收光谱法（HR-cs-GFAAS）实现了分子吸收（MAS）模式下的氟检测。然而，到目前为止，HR-cs-GFAAS还没有与分离系统（如HPLC）在线耦合，这是氟形态分析所必需的。因此，本项目的主要目标是：1)耦合界面设计：设计、构建并安装用于纳米高效液相色谱与HR-cs-GFMAS在线耦合的微液滴发生器。此外，还将解决石墨炉加热程序对瞬态信号的适应和优化、原始数据的提取和数据评价等问题。二)氟形态新开发的氟形态分析界面参数将采用氟化的测试物质进行优化。此外，将开发一种纳米高效液相色谱分离几种含氟有机测试物质的方法，以研究GFMAS在有机溶剂梯度依赖性中的种非特异性响应。开发的系统将在LOD， LOQ，线性，鲁棒性，日内和日内重现性方面进行验证。III)实际样品分析将开发和优化水样制备以及针对预期的低环境浓度氟化有机物和相关物种的样品/物种富集策略。通过纳米高效液相色谱/HR-cs-GFMAS和互补纳米高效液相色谱/ESI-Q-ToF-MS对废水处理厂和地表水的水样进行采样和分析，以揭示检测到的氟物种的分子结构。IV)适合常规分析计划简化新开发的界面，以获得一种适用的氟形态形成方法，可转移到其他实验室以及进一步的应用领域。

In the field of pharmaceuticals fluorination of organic molecules leads to new, promising properties (e.g. changed metabolism of drugs). Fluorinated substances are also beneficial in the area of technical applications.Growing production volumes and application fields lead to an increase of these substances within the (aquatic) environment. Due to their new properties a different distribution, speciation as well as bioavailability in the environment is most likely.Analytical techniques comprising high sensitivity, fluorine-selectivity as well as a fluorine species-unspecific detector response are missing. Thus, the main objective of this project is the development of a complementary method, which enables fluorine-speciation analysis for various applications including analysis of (aquatic) environmental samples to assess the load situation, distribution and bioavailability as well as speciation of fluorinated compounds. Further fields that would profit from such method are pharmaceutical, medical as well as material sciences.High resolution-continuum source -graphite furnace atomic absorption spectrometry (HR-cs-GFAAS) enables fluorine detection in the molecular absorption (MAS) mode. However, up to now HR-cs-GFAAS has not been coupled on-line with a separation systems (e.g. HPLC), which is mandatory for fluorine speciation analysis.Thus, the main objectives of this project are:I) Coupling-interface designA new interface comprising a micro-droplet generator for the on-line coupling of nanoHPLC with HR-cs-GFMAS will be designed, constructed and installed. In addition the adaption and optimization of the graphite furnace heating program for transient signals as well as raw data extraction and data evaluation will be tackled.II) Fluorine SpeciationThe parameters of the newly developed interface for fluorine speciation analysis will be optimized with fluorinated test substances. Furthermore, a nanoHPLC separation method for several fluorine-containing organic test substances will be developed to investigate the species-unspecific response of GFMAS in dependence of organic solvent gradients. The developed system will be validated in terms of LOD, LOQ, linearity, robustness, intra- & inter-day reproducibility.III) Real sample analysisWater sample preparation as well as sample/species enrichment strategies for fluorinated organic substances and related species with regard to expected low environmental concentrations will be developed and optimized. Water samples from waste water treatment plants as well as surface waters will be sampled and analyzed by means of nanoHPLC/HR-cs-GFMAS and complementary nanoHPLC/ESI-Q-ToF-MS to unravel molecular structures of the detected fluorine species.IV) Suitability for routine analysisSimplification of the newly developed interface is planned to obtain an applicable method for fluorine speciation which is transferable to other laboratories as well as further fields of application.