Rapid Identification of Active Agents and Metabolomics of Botanical Supplements

RFA-AT-11-001；CAM天然产物的机理研究(R01) 植物补充剂活性物质的快速鉴定及代谢组学 摘要： 该应用程序满足了对高吞吐量方法以识别活动组件的未满足需求 植物内膳食补充剂并通过代谢组学研究确定其作用机制。 本实验室发明的超滤质谱(PUF-MS)比传统的超滤质谱更高效。 用于筛选、表征、分离和鉴定的常规生物测定导向分级法 植物性膳食补充剂或其他天然产品中含有的具有药理意义的配体 混合物。我们建议使用一种新的超高速PUF-MS将这项技术的生产率提高100倍 (UPUF-MS)方法，并为植物膳食标准化开发新的有效分析方法 基于UHPLC-MS-MS的补充剂比以前可能的速度至少快10倍。我们会 将UHPLC-MS用于临床试验存档的血清和尿样的代谢组学研究 植物膳食补充剂黑升麻、红三叶草、啤酒花、普乐普和安慰剂对热的影响 更年期女性的闪光。我们的假设是，这种代谢组学方法将促进这一发现 指示血管运动症状的生物标记物，如潮热和盗汗。然后，这些生物标志物 可用于客观、定量地测定植物膳食补充剂对热的影响。 闪光，而不是依赖于临床试验中受试者对潮热的主观自我报告。总的来说，这些 研究将解决植物性膳食补充剂含有药理活性这一中心假设。 可使用UPUF-MS鉴定的化合物，用于UHPLC-MS-MS的化学标准化和 这些活性成分的体内效应可以用UHPLC-MS-MS联合检测 代谢组学。

RFA-AT-11-001; Mechanistic Research on CAM Natural Products (R01) Title: Rapid Identification of Active Agents and Metabolomics of Botanical Supplements Summary: This application addresses the unmet need for high-throughput methods to identify active components inbotanical dietary supplements and to determine their mechanisms of action through metabolomics studies. Ultrafiltration mass spectrometry (PUF-MS), which was invented in this laboratory, is more efficient than conventional bioassay-guided fractionation for the screening, characterization, isolation, and identification of pharmacologically significant ligands contained in botanical dietary supplements or other natural product mixtures. We propose to enhance the productivity of this technology 100-fold using a new ultrafast PUF-MS (UPUF-MS) approach and to develop new validated assays for the standardization of botanical dietary supplements based on UHPLC-MS-MS that are at least 10-fold faster than previously possible. We will incorporate UHPLC-MS into metabolomics studies of archived serum and urine samples from clinical trials of the effects of botanical dietary supplements black cohosh, red clover, hops, Prempro, and placebo on hot flashes in menopausal women. It is our hypothesis that this metabolomics approach will facilitate the discovery of biomarkers indicative of vasomotor symptoms such as hot flashes and night sweats. Then, these biomarkers may be used for the objective, quantitative determination of the effects of botanical dietary supplements on hot flashes instead of relying on subjective self reporting of hot flashes by subjects in clinical trials. Overall, these studies will address the central hypothesis that botanical dietary supplements contain pharmacologically active compounds that can be identified using UPUF-MS, used for chemical standardization with UHPLC-MS-MS and that the in vivo effects of these active constituents can be detected using UHPLC-MS-MS in combination with metabolomics.