This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).
With this award from the Major Research and Instrumentation (MRI) program, Kevin P. Minbiole and colleagues Daniel M. Downey, Reid N. Harris, Gina M. MacDonald and Debra L. Mohler of James Madison University will acquire a liquid chromatograph interfaced mass spectrometer (LC-MS). The instrument will be used in undergraduate research activities and in advanced undergraduate chemistry classes. Projects to be investigated include characterization of anti-fungal metabolites from the skin bacteria of frogs and salamanders, determination of endocrine receptor compounds in the Shenandoah River watershed, measurement of neurotransmitter metabolites in blood, and studies of the interactions between DNA and chromatin. Collaborations with Gallaudet University and Mary Baldwin College will allow deaf and hard of hearing students as well as substantial numbers of women to participate in important research endeavors.
A LC-MS combines the physical separation capability of liquid chromatography with the mass analysis ability of mass spectrometry. The liquid chromatograph separates a mixture into its molecular components. These components flow into a mass spectrometer where their masses, and those of their fragments, are measured. This highly sensitive technique allows detection and determination of the structure of molecules in a complex mixture. The instrumentation provided by this award will provide faculty and undergraduate students the opportunity to pursue significant collaborative investigations at the interface of chemistry and biology.
This award, in conjunction with NSF MRI award 1046630, is responsible for the establishment of the Undergraduate LC/MS Research & Teaching Laboratory at James Madison University. The lab houses three Agilent LC/MS instruments: a triple quadrupole (QqQ) mass spectrometer, a time of flight (TOF) mass spectrometer and a quadrupole time of flight mass spectrometer (q-TOF). These instruments are coupled to liquid chromatographs (LCs) to facilitate the analysis of complex mixtures. In addition, an on-going collaboration with Agilent Technologies ensures that the latest data acquisition and data analysis software is available for use by JMU students and faculty. The lab significantly enhances interdisciplinary teaching and research opportunities at JMU. Since its installation in 2010, seven faculty, 13 undergraduate research students and over 100 Instrumental Analysis students have used the lab. Projects are varied and include: Plant proteomics Studies of negative ion electrospray ionization efficiency Identification and quantitation of anti-fugal metabolites on the skin bacteria of frogs and salamanders Quantitation of neurotransmitter metabolites in canine urine Exact mass measurements of synthetic products Characterization of natural products, such as almonds, cranberries, cocoa, pomegranate and grapes Food metabolomics Research in these areas has lead to publications with undergraduate co-authors in peer-reviewed journals, such as Analytical Chemistry, Journal of Chemical Ecology, Food Chemistry and the Journal of Agriculture and Food Chemistry. Currently, articles are in preparation for the Journal of the American Society for Mass Spectrometry, Bioorganic and Medical Chemistry Letters and Journal of Medicinal Chemistry. Below are a couple of highlights taken from our published work that we think will be of greatest interest to the general public. Identification and quantitation of anti-fugal metabolites on the skin bacteria of frogs and salamanders A powerful mechanism for protection against disease in amphibians is synergy between metabolites produced by bacteria on the frog’s skin and antimicrobial peptides (AMPs) produced by the frog itself. We studied this method of protection in frogs (Rana muscosa) in regard to the lethal disease chytridiomycosis, which is caused by Batrachochytrium dendrobatidis (Bd). In this study, we show that the AMPs produced by Rana muscosa, as well as the metabolite 2,4-DAPG from a bacterial species normally found on the skin of R. muscosa, were inhibitory to the growth of Bd in vitro. These results may aid conversation efforts to augment amphibian skin by introducing anti-Bd species that work synergistically with amphibians’ AMPs in order to enhance resistance to chytridiomycosis. ©Journal of Chemical Ecology, p. 958, vol. 38, (2012). Characterization of natural products, such as almonds, cranberries, cocoa, pomegranate and grapes Proanthocyanidins (PACs) and ellagitannins, referred to as "tannins", occur in many plant sources. We hypothesized that tannins could bind to the digestive enzymes such as α-amylase and glucoamylase. These enzymes break starches into simple sugars, which can result in blood sugar spikes. Slowed starch digestion can theoretically increase satiety by modulating glucose "spiking" and depletion that occurs after carbohydrate-rich meals. In our study tannins were isolated from extracts of pomegranate, cranberry, grape, and cocoa and these isolates tested for effectiveness of α-amylase and glucoamylase inhibition in vitro. Results showed inhibition of each enzyme by each tannin, but with variation in magnitude. Generally, larger and more complex tannins, such as those in pomegranate and cranberry, more effectively inhibited the enzymes than did less polymerized cocoa tannins. ©Journal of Agriculture and Food Chemistry, p. 1477, vol. 61, (2013).
