This application addresses the unmet need for high-throughput methods to identify active components in botanical 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 te 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.

Public Health Relevance

Botanical dietary supplements are used by a significant portion of the population in the world including the United States. Although chemical and biological standardization of these supplements would enhance their safety and benefits, their active constituents and mechanisms of action, which are often unknown, must be determined first. This application will develop faster and more efficient methods to determine the mechanisms of action of botanical dietary supplements and to identify their active compounds. Then, faster analytical assays will be developed to support the standardization of botanical dietary supplements.

National Institute of Health (NIH)
National Center for Complementary & Alternative Medicine (NCCAM)
Research Project (R01)
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Special Emphasis Panel (ZAT1-SM (27))
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Hopp, Craig
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University of Illinois at Chicago
Schools of Pharmacy
United States
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Rush, Michael D; Rue, Emily A; Wong, Alan et al. (2018) Rapid Determination of Procyanidins Using MALDI-ToF/ToF Mass Spectrometry. J Agric Food Chem 66:11355-11361
Huang, Lingyi; Nikolic, Dejan; van Breemen, Richard B (2017) Hepatic metabolism of licochalcone A, a potential chemopreventive chalcone from licorice (Glycyrrhiza inflata), determined using liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 409:6937-6948
Rush, Michael D; Walker, Elisabeth M; Prehna, Gerd et al. (2017) Development of a Magnetic Microbead Affinity Selection Screen (MagMASS) Using Mass Spectrometry for Ligands to the Retinoid X Receptor-?. J Am Soc Mass Spectrom 28:479-485
Li, Guannan; Simmler, Charlotte; Chen, Luying et al. (2017) Cytochrome P450 inhibition by three licorice species and fourteen licorice constituents. Eur J Pharm Sci 109:182-190
Li, Guannan; Nikolic, Dejan; van Breemen, Richard B (2016) Identification and Chemical Standardization of Licorice Raw Materials and Dietary Supplements Using UHPLC-MS/MS. J Agric Food Chem :
Rush, Michael D; Walker, Elisabeth M; Burton, Tristesse et al. (2016) Magnetic Microbead Affinity Selection Screening (MagMASS) of Botanical Extracts for Inhibitors of 15-Lipoxygenase. J Nat Prod 79:2898-2902
Li, Guannan; Huang, Ke; Nikolic, Dejan et al. (2015) High-Throughput Cytochrome P450 Cocktail Inhibition Assay for Assessing Drug-Drug and Drug-Botanical Interactions. Drug Metab Dispos 43:1670-8
Huang, Ke; Huang, Lingyi; van Breemen, Richard B (2015) Detection of reactive metabolites using isotope-labeled glutathione trapping and simultaneous neutral loss and precursor ion scanning with ultra-high-pressure liquid chromatography triple quadruple mass spectrometry. Anal Chem 87:3646-54
Hajirahimkhan, Atieh; Simmler, Charlotte; Dong, Huali et al. (2015) Induction of NAD(P)H:Quinone Oxidoreductase 1 (NQO1) by Glycyrrhiza Species Used for Women's Health: Differential Effects of the Michael Acceptors Isoliquiritigenin and Licochalcone A. Chem Res Toxicol 28:2130-41