CORE--LIQUID CHROMATOGRAPHY/TANDEM MASS SPECTROMETRY

核心——液相色谱/串联质谱

• 批准号: 6300330
• 负责人: RICHARD B VAN BREEMEN
• 金额: $ 17.72万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2001-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6300330
• 关键词: alternative medicine; antioxidants; bioassay; biomedical facility; cancer prevention; carcinogenesis inhibitor; chemical structure; chemical synthesis; chemoprevention; drug discovery /isolation; liquid chromatography; mass spectrometry; oxidative stress; plant extracts; prostaglandin endoperoxide synthase; ultrafiltration

The LC-MS-MS Core will provide analytical support using liquid chromatography-tandem mass spectroscopy (LC-MS-MS) for all projects within this chemoprevention program. LC-MS-MS will be used to characterize active natural products contained in biologically active plant extracts with respect to molecular weight, chromatographic retention time, UV/vis absorbance (with in-line absorbance detection) and structure (based on MS-MS fragmentation patterns). These data will contribute to the structural elucidation of novel active compounds as well as identification of known compounds (a process known as dereplication) in support of Project 2. Because these analyses are fast (typically less than 1 hour each), they may be used to provide feedback to guide the compound scale-up activities. In addition, the purity, molecular weight, and structure of synthetic reference compounds and intermediates prepared will be confirmed by using LC-MS-MS. Whether providing data for the structural elucidating of new active natural products, identification of known natural products, or confirmation of the structures of synthetic compounds, LC-MS-MS will significantly improve the efficiency of several projects. Another function of the LC-MS-MS Core will be to contribute to the bioassay efforts by developing and carrying out screening assays using pulsed ultrafiltration mass spectrometry, which is a screening method invented in this laboratory. Specifically, chemopreventive natural products contained in plant extracts will be identified using pulsed ultrafiltration mass spectrometry based on their binding to cyclooxygenase. Once identified, the affinity constants of these natural products may be determined using pulsed ultrafiltration. Finally, the LC-MS-MS Core will interact with Projects by supporting studies on the mechanism of action of lead chemoprevention agents. For example, an LC-MS-MS assay developed in this laboratory will be used to quantitatively determine the antioxidant activity of lead compounds by measuring their ability to prevent oxidation of specific DNA bases. Since DNA oxidation might result in carcinogenesis or tumor promotion, the prevention of oxidative stress by natural products is a potential mechanism of chemoprevention. In summary, the LC-MS-MS Core will interact will all five Projects within this program by quickly providing essential mass spectroscopic data on active natural products and, as a result, this core facility will significantly enhance the overall efficiency of the program.

LC-MS-MS核心将使用该化学预防计划中所有项目的液相色谱量表质谱（LC-MS-MS）提供分析支持。 LC-MS-MS将用于表征有关分子量，色谱保留时间，UV/VIS吸光度（具有在线吸光度检测）和结构（基于MS-MS片段化）的生物活性植物提取物中包含的活性天然产物模式）。这些数据将有助于新型活性化合物的结构阐明，并识别已知化合物（一种称为删除的过程）以支持项目2。提供反馈以指导复合规模的活动。另外，通过使用LC-MS-MS确认制备的合成参考化合物和中间体的纯度，分子量和结构。无论是提供新活跃天然产品的结构性阐明，已知天然产物的识别，还是确认合成化合物的结构，LC-MS-MS将显着提高多个项目的效率。 LC-MS-MS核心的另一个功能是通过使用脉冲超滤质谱法开发和进行筛选测定法为生物测定的工作做出贡献，这是本实验室中发明的一种筛选方法。具体而言，将使用基于脉冲超滤质谱法根据其与环氧合酶的结合来鉴定植物提取物中含有的化学预防天然产物。一旦确定，这些天然产物的亲和力常数可以使用脉冲超滤确定。最后，LC-MS-MS核心将通过支持有关铅化学预防剂作用机理的研究与项目相互作用。例如，该实验室中开发的LC-MS-MS分析将用于定量确定铅化合物的抗氧化活性，通过测量其防止特定DNA碱基的氧化能力。由于DNA氧化可能导致癌变或肿瘤促进，因此预防天然产物氧化应激是化学预防的潜在机制。总而言之，LC-MS-MS Core将通过快速提供有关活动天然产品的基本质谱数据，将在该计划中进行交互，因此，该核心设施将显着提高该计划的整体效率。