2010 Annual Report
Objective 2: Develop and evaluate spectral fingerprinting and chromatographic profiling methods to characterize components in lipid soluble, water soluble, and intermediate fractions of food materials. Sub-objective 2.A.: Develop spectral fingerprinting methods for identification of plant materials and individual components using direct analysis (no prior chromatographic separation) and pattern recognition algorithms. Sub-objective 2.B.: Develop chromatographic profiling methods for identification and quantification of individual components in plant materials.
Objective 3: Develop methods to determine variability of biologically active components in food materials through profiles and/or fingerprints.
Objective 2: Spectral fingerprinting methods will be developed based on ultraviolet and visible molecular absorption (UV/Vis), infrared (IR), near-infrared (NIR), and mass spectrometric (MS) detection. The overlapping complex spectra will be interpreted using pattern recognition programs. The patterns will be used to determine the sensitivity of the different detection systems for discriminating between plant materials based on genera, species, variety, growing year, growing site, and processing conditions. These methods will be developed using 3 food materials and 3 botanical supplement materials. Repeat samples will be examined over a period of years to determine the stability of the spectra and the ability to compare spectra of new materials to archived spectra. The phenolic and vitamin content of the plant materials will also be determined using chromatographic profiling using LC-DAD-ESI/MS. This will make it possible to determine which compounds are contributing most to differences arising from the various growing factors.
Objective 3: The spectral fingerprints can be used with nested analysis of variance to determine the relative variance contributed by each growing factor: species, variety, site, year, plant-to-plant variation, and analytical uncertainty. Samples will be obtained from collaborators across the country and representing a variety of foods and botanical supplements. UV/Vis, IR, and NIR spectra will provide variance data for the integrated chemical composition of the plant materials and MS will provide variance data for specific masses and, with the assistance of chromatographic profiling, specific compounds of health interest.
An ASE method was optimized for Artemisinin, an anti-viral, anti-carcinogenic, sesquiterpene lactone found in Artemesia, that is of medical interest. We optimized our method to investigate the effect of drying procedures and growing conditions (field, greenhouse, or tissue culture) on artemisinin, dihydroartemisinic acid (a precursor), and the phenolic content of the tissue. This work was in collaboration with Applachian Farming Systems Research Center (ARS, USDA, Beaver, WV).
FCMDL continued development of methods using spectral fingerprinting and pattern recognition programs to characterize botanicals. In 2010 we examined 3 ginseng species: American (Panax quinquefolius), Chinese ginseng (P. quinquefolius), and P. notoginseng. This project was supported by the Office of Dietary Supplements at NIH and is described in more detail in project report 1235-52000-060-11R. Direct analysis of solids or extracts by ultraviolet (UV), near-infrared (NIR), or mass spectrometry (MS) readily identified the 3 species and the “red” and “white” preparations of the “Chinese” ginseng. Mass spectrometry (MS) and near-infrared (NIR) could also be used to identify the growing location (China, U.S., or Canada) for American ginseng. Computer programs used for pattern recognition were developed in collaboration with Ohio University (project report 1235-52000-060-08G). Using statistical analysis programs we were able to locate spectral regions (by UV and NIR) or masses (by MS) that were significantly different for the species and provided enhanced ability to distinguish between them.
The standardized method for analysis of polyphenols (developed at FCMDL) was used to analyze flavonoids and phenolic acids in Brassica olaracea (e.g. kale, collard greens, and broccolis) and other Brassica species (e.g. mustard greens, bok choy, and napa). More than 100 phenolic compounds were identified in both groups, many for the first time. A method is being developed for the quantification of glycosylated flavonoids based on general rules for shifts in the peak wavelengths and absorption coefficients for the conjugated forms.
Lin, L., Harnly, J.M. 2010. Identification of phenolic components of chrysanthemum flower (Chrysanthemum morifolium Ramat). Food Chemistry 120(1):319-326.
Singh, A.P., Luthria, D., Wilson, T., Vorsa, N., Singh, V., Banuelos, G.S., Pasakdee, S. 2009. Polyphenol content and antioxidant capacity of eggplant pulp. Food Chemistry. 114(3):955-961.
Chen, P., Song, F., Lin, L. 2009. Chromatographic fingerprint analysis of Pycnogenol® dietary supplements. Journal of AOAC International. 92(2):624-632.
Tansupo, P., Suwannasom, P., Luthria, D.L., Chanthai, S., Ruangviriyachai, C. 2010. Optimized separation procedures for the simultaneous assay of three plant hormones in liquid biofertilizers. Phytochemical Analysis. 21(2):157-162.
Memon, A.A., Memon, N., Luthria, D.L., Bhanger, M.I., Pitafi, A.A. 2010. Phenolic acids profiling and antioxidant potential of mulberry (Morus laevigata W., Morus nigra L., Morus alba L.) leaves and fruits grown in Pakistan. Polish Journal of Food and Nutrition Sciences. 60(1)25-32.
Luthria, D., Singh, A.P., Wilson, T., Vorsa, N., Banuelos, G.S., Vinyard, B.T. 2010. Influence of conventional and organic agricultural practices on the phenolic content in eggplant pulp: Plant-to-plant variation. Food Chemistry. 121(2):406-411.
Chen, P., Lin, L., Harnly, J.M. 2010. Mass spectroscopic fingerprinting method for differentiation between Scutellaria lateriflora and the Germander (Teucrium canadense and T. chamaedrys) species. Journal of AOAC International. 93(4):1148-1154.
Chen, P., Harnly, J.M., Lester, G.E. 2010. Flow injection mass spectral fingerprints demonstrate chemical differences in Rio Red grapefruit with respect to year, harvest time, and conventional versus organic farming. Journal of Agriculture and Food Chemistry. 58(8):4545-4553.
Harnly, J.M., Lin, L. 2010. Phenolic component profiles of mustard greens, yu choy and 15 other Brassica vegatables. Journal of Agricultural and Food Chemistry. 58(11):6850-6857.
Luthria, D.L. 2009. Phenolic compounds analysis in foods and dietary supplements is not the same using different sample preparation procedures. Acta Horticulturae Proceedings (ISHS). 841:381-388.