CORE--ANALYTICAL

分析核心将提供LC-MS（液相色谱-质谱法）、LC-MS-MS（液相色谱-串联质谱法）和MS-MS（串联质谱法）分析，以支持Farnsworth、货车Breeman和博尔顿在该植物中心开展的项目。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。这一过程的一部分将包括去复制，即快速鉴定先前报告的植物中存在的化合物，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，以及货车Breemer项目中支持啤酒花I期临床试验的血清和尿液样本。在特定目标2中，将使用超滤LC-MS（本实验室发明的一种技术）筛选植物提取物，以快速鉴定雌激素受体和5-羟色胺受体的配体。这些高 通量筛选分析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，将基于1，1-二苯基-2-苦肼（DPPH）自由基清除试验、作为脂质过氧化指示剂的丙二醛测定和作为氧化应激生物标志物的花生四烯酸非酶分解产物的8-isoPGF 2a测定，开发LC-MS和LC-MS-MS测定。这些测定将补充博尔顿项目的抗氧化剂目标3和货车Breemer项目的抗氧化剂目标2，从而有助于提供正在研究的植物膳食补充剂的抗氧化剂特性的广泛情况。最后，具体目标4将使用人肠道吸收的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，以支持用于女性健康的活性植物膳食补充剂的临床前开发。正如该实验室与上一个资助期内所有四个项目之间的协同作用所示，该核心实验室将继续提高整个植物中心的生产力。

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.