Rapid Identification of Active Agents and Metabolomics of Botanical Supplements

RFA-AT-11-001; CAM天然产物的机理研究（R 01） 标题：植物性补充剂的活性剂和代谢组学的快速鉴定 总结： 该应用解决了高通量方法识别活性组分的未满足需求 我们希望通过代谢组学研究来确定它们的作用机制。 本实验室发明的超滤质谱法（PUF-MS）比 传统的生物测定引导的分级分离，用于筛选、表征、分离和鉴定 植物膳食补充剂或其他天然产物中含有的非显著配体 混合物。我们建议使用新的超快PUF-MS将该技术的生产率提高100倍 （UPUF-MS）方法，并开发新的经验证的测定方法，用于植物膳食标准化 基于UHPLC-MS-MS的补充剂比以前可能的速度快至少10倍。我们将 将UHPLC-MS纳入临床试验存档血清和尿液样本的代谢组学研究， 植物性膳食补充剂黑升麻、红三叶草、啤酒花、Prempro和安慰剂对热的影响 更年期妇女的闪光我们的假设是，这种代谢组学方法将有助于发现 指示血管炎症状的生物标志物，如潮热和盗汗。然后，这些生物标志物 可用于客观，定量测定植物膳食补充剂对热的影响， 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 研究将解决植物膳食补充剂含有抗氧化剂的中心假设， 可以使用UPUF-MS识别的化合物，用于使用UHPLC-MS-MS进行化学标准化，以及 这些活性成分的体内作用可以使用UHPLC-MS-MS结合 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.