Analytical Chemistry

分析化学

• 批准号: 9551326
• 负责人: Anton Simeonov
• 金额: $ 194.8万
• 依托单位: NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9551326
• 关键词: Adopted; Adoption; Analytical Chemistry; Binding Proteins; Biochemical; Biological Assay; Chemicals; Chemistry; Chromatography; Collaborations; Collection; Complement; Computer software; Data; Data Analyses; Data Storage and Retrieval; Development; Eligibility Determination; Ensure; Equipment and supply inventories; Extramural Activities; Fluorescence; Goals; High Pressure Liquid Chromatography; Industry Standard; Informatics; Journals; Laboratories; Libraries; Liquid substance; Mass Fragmentography; Mass Spectrum Analysis; Methods; Minor; Mission; Molecular Conformation; Molecular Models; Monitor; NMR Spectroscopy; Nuclear Magnetic Resonance; Optical Rotation; Paper; Peer Review; Peptides; Pharmaceutical Chemistry; Phase; Preparation; Process; Productivity; Protein Analysis; Proteomics; Protocols documentation; Reaction; Reagent; Research; Resolution; Resources; Sampling; Scientist; Synthesis Chemistry; System; Techniques; Testing; Therapeutic; Time; Tube; Vial device; Vibrational Circular Dichroism; Work; Writing; assay development; base; data management; detector; high throughput screening; improved; innovation; instrumentation; light scattering; liquid chromatography mass spectrometry; metabolomics; method development; milligram; molecular modeling; multiple reaction monitoring; new technology; pre-clinical; programs; scale up; screening; small molecule; ultraviolet

The Analytical Chemistry team has continued to perform the core responsibility of purifying samples with material in the range of milligrams to grams. Major and minor components (< 0.1 percent) have been isolated for additional testing and characterization. The teams automated sample processing protocol allows for dispensing into 1D barcoded vials, Matrix 2D barcoded tubes and/or 96-well plates for efficient tracking, storing and testing. The entire process from receiving the sample to final plating is completed within one week. While the group uses a variety of liquid chromatographs to determine identity and purity, single quadrupole liquid chromatography/mass spectrometry instrumentation is utilized for high-throughput automated analysis. Due to the wide variety of analytes tested, the teams range of analytical detectors includes ultraviolet (UV), mass spectrometry (MS; positive and negative mode), and evaporative light scattering detector and fluorescence (ELSD). Time-of-flight mass spectrometry (TOF/MS) is employed to achieve formula confirmation and identity determination of unknowns. The chemical and enantiomeric purity of chiral compounds is routinely determined within the groups full scale chiral laboratory. Methods development with the chiral chromatography screening protocol involves the utilization of various chiral stationary phases in conjunction with multiple mobile phase conditions. The use of an inline chiral detector allows for the determination of relative optical rotation. Sample purification on a scale of up to hundreds of milligrams is possible. In an effort to analyze and separate any chiral compound type, we have expanded our capabilities by increasing the number of different chiral columns at our disposal over the past year. Additionally, utilization of our vibrational circular dichroism (VCD) spectrometer has allowed for determining the absolute configuration of isolated chiral compounds. This analysis is a collaborative effort with the DPI Informatics group to utilize their unparalleled molecular modeling capabilities. The Sample Management and Resource Tracking (SMART) system, our proprietary LIMS, continues to be utilized at DPI for sample submission, compound purification, compound processing, sample registration, compound inventory, sample tracking, data retrieval, and data management. As SMART is constantly evolving based on changing workflows and scientist needs, we have add required functionality while also streamlining user interfaces and the underlying scripting, which has led to a more robust and efficient system. In collaboration with NCATS OIT, we are in the process of transitioning SMART to a unified platform, which will enable better system control and higher integration with other software at DPI. Additionally, the SMART Consumables, Acquisitions, and Reagent Tracker (SMARTCART), which aids in the requisition, organization, and management of scientific related orders has be highly successful within DPI Chemistry. As such, the NCATS Acquisitions Workgroup has evaluated SMARTCART and we are working towards eventual adoption by all DPI scientific groups. Efforts continue in the establishment of our NMR fragment-based screening (FBS) program, which is a complementary approach to high-throughput screening in the discovery of chemical entities for development into therapeutics. With the integration of the fragment libraries complete, the team has begun the massive task of compiling reference spectra for each fragment within the collection. Analytical Chemistry maintains several collaborations conducting NMR ligand-protein binding studies as a complement to the established medicinal chemistry programs. Furthermore, we are expanding our capabilities to employ data fitting of multiple NMR parameters to perform configurational and conformational analysis of small molecules. Finally, we acquired a benchtop NMR for use within the Medicinal Chemistry labs. This system compliments our state-of-the-art NMR facilities by allowing for simple and expedient reaction monitoring to ensure formation of the desired product. With the successful development and set up of our state-of-the-art Mass Spectrometry laboratory, we have continued the expansion of analytical capabilities at NCATS. These include high-throughput MS sample analysis, peptide and protein analysis, multiple reaction monitoring, proteomics, and metabolomics. We have established collaborations for numerous ongoing projects at DPI with our involvement in assay development, methods optimization, mass spectroscopy experimentation, and data analysis. Utilization of the RapidFire high throughput sampling system provides us with a potential analysis capacity of 5000-10,000 samples per day (qualitative and quantitative) within various biochemical and ADME assays. Analytical Chemistry at DPI has access to thirteen mass spectrometry systems, which includes one GC-MS, eight HPLC-single Quad MS, one triple quad (QQQ) MS and three high resolution MS (two TOFs and one QTOF). We are currently writing papers for submission to peer-reviewed journals in order to highlight this extensive and high impact work.

分析化学团队继续履行纯化毫克至克范围内材料样品的核心职责。主要和次要成分（< 0.1%）已被隔离以进行额外的测试和表征。该团队的自动化样品处理协议允许分配到 1D 条形码小瓶、Matrix 2D 条形码管和/或 96 孔板中，以便高效跟踪、存储和测试。从收到样品到最终电镀的整个过程在一周内完成。虽然该小组使用各种液相色谱仪来确定身份和纯度，但单四极杆液相色谱/质谱仪器用于高通量自动分析。由于测试的分析物种类繁多，该团队的分析检测器范围包括紫外 (UV)、质谱（MS；正负模式）以及蒸发光散射检测器和荧光 (ELSD)。采用飞行时间质谱（TOF/MS）来实现化学式确认和未知物的身份确定。 手性化合物的化学和对映体纯度通常在集团全规模手性实验室内测定。手性色谱筛选方案的方法开发涉及各种手性固定相与多种流动相条件的结合。使用在线手性检测器可以确定相对旋光度。样品纯化规模可达数百毫克。为了分析和分离任何手性化合物类型，我们在过去一年中通过增加可用的不同手性色谱柱的数量来扩展我们的能力。此外，利用我们的振动圆二色性 (VCD) 光谱仪可以确定分离的手性化合物的绝对构型。该分析是与 DPI 信息学小组合作进行的，旨在利用他们无与伦比的分子建模能力。 样品管理和资源跟踪 (SMART) 系统（我们专有的 LIMS）继续在 DPI 中用于样品提交、化合物纯化、化合物处理、样品注册、化合物库存、样品跟踪、数据检索和数据管理。由于 SMART 根据不断变化的工作流程和科学家需求不断发展，我们添加了所需的功能，同时简化了用户界面和底层脚本，从而形成了更加强大和高效的系统。我们正在与 NCATS OIT 合作，将 SMART 过渡到统一平台，这将实现更好的系统控制以及与 DPI 的其他软件的更高集成。此外，有助于申请、组织和管理科学相关订单的 SMART 耗材、采购和试剂跟踪器 (SMARTCART) 在 DPI Chemistry 中取得了巨大成功。因此，NCATS 采集工作组已经评估了 SMARTCART，我们正在努力最终被所有 DPI 科学小组采用。 我们继续努力建立基于 NMR 片段的筛选 (FBS) 计划，该计划是在发现化学实体以开发成治疗药物方面对高通量筛选的补充方法。随着片段库的集成完成，该团队开始了为集合中的每个片段编译参考光谱的艰巨任务。分析化学保持着多项合作，进行核磁共振配体-蛋白质结合研究，作为对已建立的药物化学项目的补充。此外，我们正在扩展我们的能力，利用多个 NMR 参数的数据拟合来进行小分子的构型和构象分析。最后，我们购买了一台台式核磁共振仪供药物化学实验室使用。该系统通过允许简单而方便的反应监测来确保所需产物的形成，从而补充了我们最先进的 NMR 设施。 随着我们最先进的质谱实验室的成功开发和建立，我们继续扩展 NCATS 的分析能力。其中包括高通量 MS 样品分析、肽和蛋白质分析、多反应监测、蛋白质组学和代谢组学。我们与 DPI 的众多正在进行的项目建立了合作关系，参与分析开发、方法优化、质谱实验和数据分析。 RapidFire 高通量采样系统的利用为我们提供了在各种生化和 ADME 测定中每天 5000-10,000 个样本（定性和定量）的潜在分析能力。 DPI 分析化学部门拥有 13 套质谱系统，其中包括一套 GC-MS、八套 HPLC-单四极杆 MS、一套三重四极杆 (QQQ) MS 和三套高分辨率 MS（两个 TOF 和一套 QTOF）。我们目前正在撰写论文以提交给同行评审期刊，以突出这项广泛且具有高影响力的工作。