With this award from the Major Research Instrumentation (MRI) program, Professor Elizabeth Stemmler from Bowdoin College and colleagues Patsy Dickinson, Dharni Vasudevan, Danielle Dube and Benjamin Gorske will acquire a liquid chromatograph-quadrupole, mass spectrometer with tandem capabilities and various accessories. The proposal will enhance research training and education at all levels, especially in areas such as (a) use of transcriptomics and peptidomics strategies to neuropeptide identification and to explore the role of hormonal cycles on differential neuromodulatory responses in crustaceans, (b) characterization of bacterial glycosylation, (c) studies of protein methylation, (d) characterization of transformations of molecules of importance to the environmental degradation of contaminants, and (e) development of methods to synthesize and characterize oligothioamides.

Mass spectrometers (MS) are used to identify the chemical composition of a sample by measuring the mass of the molecular constituents in the sample after they are ionized and detected by the mass spectrometer. This instrument couples a high-resolution liquid chromatography system with the mass analysis ability of mass spectrometry. The liquid chromatograph separates mixtures into their molecular components. These components then flow into a mass spectrometer where their masses, and those of their fragments, are measured. This mass spectrometer has a special design that provides relatively high mass accuracy, sensitivity and resolution that allows detection and determination of the structure of molecules in a complex mixture. The instrumentation will be used not only for research but also in laboratory courses to train significant numbers of students in the use of this important analytical technique and for research and outreach activities with regional institutions such as Bates College, Colby College and Mount Desert Island Biological Laboratory.

Project Report

Normal 0 false false false EN-US X-NONE X-NONE This grant, awarded for the acquisition of an Chip-based nanoESI-LC-Q-TOF MS/MS mass spectrometer, has improved the quality and expanded the scope of primarily undergraduate research and training activities in chemistry, biology, neuroscience, and environmental science at Bowdoin College and at three additional institutions. The instrument has provided unique data in studies directed at understanding the composition and chemical transformations of wood-derived pyrolysis bio-oils, where the ability to analyze higher molecular mass components has extended the range of compounds that can be characterized in ways that have informed the modification of pyrolysis conditions and impacted the development of woody biomass as a potential bio-oil source. Our work on the detection of labelled glycans has provided validation and initiated further studies directed at the analysis of bacterial glycans released from enriched, glycosylated proteins(Champasa et al. 2013). Our work on neuropeptide extraction has revealed a pathway leading to neuropeptide modifications that are extraction artifacts, not endogenous neuropeptide modifications, which has resulted in improved methods for the extraction and analysis of neuropeptides from complex tissues (Stemmler et al. 2013); furthermore, we are developing analytical methods for shotgun proteomic analyses to characterize proteins present in crustacean nervous system tissues. Our work directed at the characterization of photodegradation products derived from estrogens and environmental contaminants has assisted researchers as they develop newer and more effective catalysts for the treatment of wastewaters (Pan et al. 2014). The acquisition of the LC-MS instrument has expanded the scope of research questions addressed by students and faculty at Bowdoin and nearby institutions. We have been able to analyze intact proteins, of benefit to co-PI-Danielle Dube and Professor Rachel Austin (Bates) and their students. We are now able to sequence and identify neuropeptides and proteins, of benefit to PI-Stemmler, co-PI-Dickinson, co-PI-Dube, and collaborators Professor Anne McBride and Andrew Christie (University of Hawaii, Manoa). We have developed techniques that have extended our ability to analyze polar components of bio-oils, in collaboration with faculty (Professors Brian Frederick, Clayton Wheeler, and William DeSisto) and graduate students (Jincy Joseph and Paige Case) at the University of Maine, Orono. Acquisition of the instrument has played a key role in confirming the identity of synthetic products produced by organic chemists, including co-PI-Gorske and his students and supporting synthetic efforts by co-PI-Dube and her students. Finally, the instrument has provided graduate students Zhong Pan and John Ahern and their faculty advisors (Professor Howard Patterson and Aria Amirbahman) at the University of Maine, Orono, access to an instrument that continues to provide key identifications of photodegradation products, supporting their research efforts at the development of photolysis catalysts. This grant has supported the research training and education of >20 undergraduates and four Ph.D. students (University of Maine, Orono), 19 of whom are women. Sixteen of these students completed year-long projects that have earned them Honors in Chemistry or Biochemistry upon graduation. The instrumentation has also been incorporated into two Bowdoin College chemistry courses (Organic Structure Determination and Instrumental Analysis), and in-house training materials and exercises have been developed and used during instrument training activities.

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Division of Chemistry (CHE)
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Carlos A. Murillo
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Bowdoin College
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