Critical Replacement of Super Critical Fluid HPLC for Chiral Separations

超临界流体 HPLC 用于手性分离的关键替代品

• 批准号: RTI-2021-00129
• 负责人: Crudden, Cathleen
• 金额: $ 10.29万
• 依托单位: Queen's University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=718803
• 关键词: Critical; Replacement; Super; Fluid; HPLC

Methods to quantify chirality are complex since enantiomers have identical physical properties. The use of optical rotation in a quantitative sense is unreliable on small scales and highly sensitive to impurities. Chiral shift reagents are not always effective, require tedious titration into a sample and are not helpful for reactions that are highly selective. Chromatography with chiral stationary phases is by far the best alternative, and the use of super critical fluids is essential to increase the speed and efficiency of separation. In our experience, compounds elute 2-5 times faster using SFC compared to traditional HPLC. Since this instrument will be shared between multiple groups, speed of analysis and development of reaction conditions is essential. A major part of the Crudden lab studies the development of enantioselective and enantiospecific cross coupling reactions. This project previously relied upon a 10 year old SFC-HPLC to separate and identify enantiomers after catalytic CC bond connections. After functioning well for 9 years, this instrument suffered a series of breakdowns, was sent back to the parent company for repair over the COVID shutdown, and is still not functioning after being returned to us. For the time being, this leaves us unable to measure chirality in our samples, which is threatening the work of an MSc student, two PhD students, and two postdoctoral fellows. Another program that will be impossible without the requested infrastructure is the use of chiral nanoclusters in catalysis. Crudden has spent the last 6 years developing the use of carbon-based NHC ligands in materials science, culminating in the successful synthesis of a series of nanoclusters, most recently describing two chiral nanoclusters. The next step is the application of these in synthesis, which will not be possible without the requested infrastructure. The Stamplecoskie group has significant experience in nanoclusters and has recently prepared a chiral Ag29 cluster, but cannot use it for anything without the requested infrastructure. Queen's is developing a strength in bio-organic chemistry and the programs of three key members of this program (Zechel, Howe and Ross) will be severely impacted without access to chiral chromatography. New hire Howe is investigating directed evolution of enzymes, including the preparation of designer enzymes to prepare chiral molecules with enzyme-level specificity. Zechel is studying iron-dependent oxygenases for use in stereospecific C-H activation, and Ross needs access to this instrument to determine stereochemistry of amino acids as part of her work in structure determination of low abundance bioactive peptides from bacterial sources. None of this work will be possible without access to the requested infrastructure, which will also serve as an excellent training ground for HQP in the five groups, and a touchpoint for interactions between HQP from these different research areas.

由于对映体具有相同的物理性质，量化手性的方法很复杂。在定量意义上使用旋光度在小尺度上不可靠并且对杂质高度敏感。手性位移试剂并不总是有效，需要对样品进行繁琐的滴定，并且对于高选择性反应没有帮助。手性固定相色谱法是迄今为止最好的替代方法，超临界流体的使用对于提高分离速度和效率至关重要。根据我们的经验，与传统 HPLC 相比，使用 SFC 的化合物洗脱速度快 2-5 倍。由于该仪器将在多个小组之间共享，因此分析和反应条件开发的速度至关重要。 克鲁登实验室的主要部分研究对映选择性和对映特异性交叉偶联反应的发展。该项目之前依靠已有 10 年历史的 SFC-HPLC 来分离和鉴定催化 CC 键连接后的对映体。该仪器在正常运行了 9 年之后，出现了一系列故障，因新冠疫情停工而被送回母公司维修，返回后仍然无法运行。目前，这使我们无法测量样本中的手性，这威胁着一名硕士生、两名博士生和两名博士后的工作。 如果没有所需的基础设施，另一个不可能实现的计划是在催化中使用手性纳米团簇。 Crudden 在过去 6 年里一直致力于开发碳基 NHC 配体在材料科学中的应用，最终成功合成了一系列纳米团簇，最近描述了两种手性纳米团簇。下一步是综合应用这些，如果没有所需的基础设施，这是不可能实现的。 Stamplecoskie 小组在纳米团簇方面拥有丰富的经验，最近制备了一个手性 Ag29 团簇，但如果没有所需的基础设施，就无法将其用于任何用途。 女王大学正在发展生物有机化学方面的优势，如果无法获得手性色谱，该项目的三个关键成员（Zechel、Howe 和 Ross）的项目将受到严重影响。新员工 Howe 正在研究酶的定向进化，包括制备设计酶以制备具有酶水平特异性的手性分子。 Zechel 正在研究用于立体特异性 C-H 激活的铁依赖性加氧酶，Ross 需要使用该仪器来确定氨基酸的立体化学，作为她对细菌来源的低丰度生物活性肽进行结构测定的工作的一部分。如果没有所需的基础设施，所有这些工作都将不可能实现，这些基础设施也将成为五个小组中总部的优秀培训基地，以及来自这些不同研究领域的总部之间互动的接触点。