Digitized Honors Theses (2002-2017)

Date of Award

5-2012

Document Type

Undergraduate Thesis

Degree Name

BS

Department

Biomedical Sciences

Faculty Mentor

Alexandra Stenson

Abstract

Humic materials are complex mixtures of decayed organic matter that can be found dissolved in water. The goal of this research was to reduce the isobaric complexity of one humic sample, Suwannee River Fulvic Acid (SRFA). SRFA was first pre­ fractionalized with reverse phase high pressure liquid chromatography (RP-HPLC). RP­ HPLC pumps a mostly polar mobile phase through a column packed with a stationary phase at high pressure. The column of choice for the pre-fractionation was a phenyl column which is packed with aromatic moieties. These aromatic moieties form pi-pi interactions with the aromatic SRFA components. Therefore, the more aromatic and nonpolar a SRFA analyte is, the later it elutes. Selected fractions from the pre­ fractionation were further fractionated with Ni2+immobilized metal affinity chromatography (IMAC) and also IMAC in sequence with an Obelisc N column to allow for further separation. To test the efficiency of these fractionations on actual humic samples, high-resolution spectra were compared through Kendrick and Van Krevelen plots, and hydrogen/deuterium exchange was performed on the SRFA fractions. Hydrogen/Deuterium exchange on SRFA reliably corresponds to the number of hydroxyl (-OH) groups..Thus, this technique allowed for functional group comparison between the different fractions.

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