Reese, R. Neil , Wyzgoski, Faith J , Miller, A. Raymond , Scheerens, Joseph C. , Rinaldi, Peter L. , Bishop, Bert , Ozgen, Mustafa , Tulio, Artemio J. , Giusti, M. Monica , Bomser, Joshua A. .
Biologically Active Plant Metabolites in Black Raspberries: A Nuclear Magnetic Resonance (NMR) Spectroscopy-Based Approach to Identify Anticancer Compounds.
A NMR-based approach to metabolomics research that enables the identification of bioactive compounds in unpurified plant extracts was developed. Black raspberries, which are known to contain compounds that exhibit chemopreventive activity toward oral, esophageal and colon cancers, were evaluated chemically and in a cancer cell bioassay. To characterize bioactive components and their interrelationships, 1H-NMR spectra of 19 black raspberry samples (four cultivars from 7 commercial berry farms in Ohio) were partitioned into 0.005 ppm segments and the areas within those fractions subjected to principal component analysis. Multivariate analysis of assay data that included anthocyanin content (HPLC), antioxidant activity (DPPH, ABTS, FRAP), total phenolics (Folin-Ciocalteau assay) and bioactivity as measured by inhibition of colon cancer HT-29 cell lines showed correlations with principle components (eigenvectors). Specific regions of 1H-NMR spectra contributing to each of the eigenvalues provide direction for identification of the specific biologically active compounds using multidimensional NMR techniques. Initial experiments were conducted with a 400 MHz spectrometer and experiments using the fully developed protocols were conducted using replicate black raspberry samples, examined with a 750 MHz NMR spectrometer, equipped with a cryoprobe that provided a 4- to5- fold improvement in sensitivity. This method utilized the natural variation in black raspberry secondary metabolites to identify biologically active chemicals and synergistic interactions between compounds without fractionation of samples before testing.
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1 - South Dakota State University, Department of Biology & Microbiology, Box 2140D, Brookings, South Dakota, 57007, USA
2 - Ohio State University, Mansfield, Department of Chemistry, Mansfield, Ohio, 44906, USA
3 - Ohio State University, Ohio Agricultural Research & Development Center, Department of Horticulture & Crop Science, Wooster, Ohio, 44691, USA
4 - University of Akron, Department of Chemistry, Akron, Ohio, 44325, USA
5 - Ohio State University, Ohio Agricultural Research & Development Center, Computing & Statistical Services, Wooster, Ohio, 44691, USA
6 - Gaziosmanpasa University, Horticulture Department, Tokat, TR-60240, Turkey
7 - Ohio State University, Department of Food Science & Technology, Columbus, Ohio, 43210, USA
8 - Ohio State University, Department of Human Nutrition, Columbus, Ohio, 43210
Presentation Type: Poster:Posters for Sections
Location: Auditorium/Bell Memorial Union
Date: Tuesday, August 1st, 2006
Time: 12:30 PM