CORE--ANALYTICAL

核心——分析

• 批准号: 6954968
• 负责人: RICHARD B VAN BREEMEN
• 金额: $ 17.2万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6954968
• 关键词: alternative medicine; biomedical facility; dietary supplements; electrospray ionization mass spectrometry; gas chromatography mass spectrometry; liquid chromatography mass spectrometry; menopause; plant extracts; premenstrual syndrome; women' s health

The Analytical Core will provide LC-MS (liquid chromatography-mass spectrometry), LC-MS-MS (liquid chromatography-tandem mass spectrometry) and MS-MS (tandem mass spectrometry) analyses to support the efforts of Projects by Farnsworth, van Breeman, and Bolton within this Botanical Center. In Specific Aim 1 the botanical natural products isolated by Project by Farnsworth will be characterized with respect to molecular weight, elemental composition (based on exact mass measurements), structure (as indicated by MS-MS fragmentation patterns), and HPLC retention time. Part of this process will include dereplication which is the rapid identification of compounds previously reported to occur in botanicals so that the efficiency of Project by Farnsworth is enhanced. In addition, quantitative LC-MS-MS will be used to measure the concentrations of active compounds in the botanical extracts in support of the standardization effort of Project by Farnsworth, and in serum and urine samples in support of the Phase I clinical trial of hops in Project by van Breemer. In Specific Aim 2 botanical extracts will be screened using ultrafiltration LC-MS (a technique invented in this laboratory) for the rapid identification of ligands to the estrogen receptors and serotonin receptors. These high throughput screening assays will complement the bioassay efforts of Project by Bolton and enhance the productivity of the entire Botanical Center. In Specific Aim 3, LC-MS and LC-MS-MS assays will be developed based on the 1,1-diphenyl-2-picrylhydrazine (DPPH) free radical scavenging assay, the measurement of malondialdehyde as an indicator of lipid peroxidation, and the measurement of 8-isoPGF2a which is a product of non-enzymatic decomposition of arachidonic acid used as a biomarker of oxidative stress. These assays will complement the antioxidant Aim 3 of Project by Bolton and Aim 2 of Project by van Breemer and will thereby help provide a broad picture of the antioxidant properties of the botanical dietary supplements under investigation. Finally, Specific Aim 4 the intestinal permeability of active botanical compounds will be assayed using the Caco-2 cell monolayer model of human intestinal absorption in combination with LC-MS and LC-MS-MS to support the preclinical development of active botanical dietary supplements for women's health. As shown by the synergy between this laboratory and all four Projects during the previous funding period, this Core laboratory will continue to enhance the productivity of the entire Botanical Center.

分析核心将提供 LC-MS（液相色谱-质谱）、LC-MS-MS（液相色谱-串联质谱）和 MS-MS（串联质谱）分析，以支持 Farnsworth、van Breeman 的项目工作和博尔顿在这个植物中心内。在具体目标 1 中，Farnsworth 项目分离的植物天然产物将在分子量、元素组成（基于精确的质量测量）、结构（如 MS-MS 碎片模式所示）和 HPLC 保留时间方面进行表征。该过程的一部分将包括去重复，即快速识别先前报道的植物药中存在的化合物，从而提高 Farnsworth 项目的效率。此外，定量 LC-MS-MS 将用于测量植物提取物中活性化合物的浓度，以支持 Farnsworth 项目的标准化工作，以及血清和尿液样本中活性化合物的浓度，以支持啤酒花的 I 期临床试验在范·布雷默的项目中。在具体目标 2 中，将使用超滤 LC-MS（本实验室发明的技术）筛选植物提取物，以快速识别雌激素受体和血清素受体的配体。这些高 通量筛选测定将补充博尔顿项目的生物测定工作，并提高整个植物中心的生产力。在具体目标 3 中，LC-MS 和 LC-MS-MS 检测将基于 1,1-二苯基-2-三硝基苯肼 (DPPH) 自由基清除检测、作为脂质过氧化指标的丙二醛测量以及8-isoPGF2a 的测量，它是花生四烯酸非酶分解的产物，用作氧化应激的生物标志物。这些测定将补充 Bolton 项目的抗氧化剂 Aim 3 和 van Breemer 项目的 Aim 2，从而有助于提供所研究的植物膳食补充剂的抗氧化特性的广泛了解。最后，具体目标 4 将使用人体肠道吸收的 Caco-2 细胞单层模型结合 LC-MS 和 LC-MS-MS 来测定活性植物化合物的肠道渗透性，以支持活性植物膳食补充剂的临床前开发女性健康。正如上一资助期间该实验室与所有四个项目之间的协同作用所表明的那样，该核心实验室将继续提高整个植物中心的生产力。