Ultra Performance Liquid Chromatography Analysis of Proteins and Synthetic Intermediates
Faculty Sponsor
Thomas Goyne
College
Arts and Sciences
Discipline(s)
Chemistry
ORCID Identifier(s)
orcid.org/0000-0003-4642-9825, orcid.org/0000-0001-8790-9089
Presentation Type
Poster Presentation
Symposium Date
Spring 4-23-2016
Abstract
The goal of this project was to develop standard operating procedures for analyzing proteins and synthetic intermediates using Ultra Performance Liquid Chromatography (UPLC), an experimental tool that is designed to separate complex mixtures into their individual components. Specifically, two classes of molecules were subjected to UPLC analysis: 1) proteins and 2) intermediates in the synthesis of fluorescent nonnatural amino acids. Proteins were analyzed following tryptic digest (i.e, after breaking the protein into well-defined fragments by the action of the digestive enzyme trypsin) using mass spectroscopy (MS) detection. In many cases, the results allow the definitive identification of the parent protein. The analysis of synthetic intermediates was focused on the synthesis of nonnatural amino acids. Three types of detection were used in the analysis of synthetic intermediates from the preparation of non-natural fluorescent amino acids: ultraviolet-visible (UV-vis), mass spectroscopy (MS), and fluorescence. The UPLC results provided information that complemented results of nuclear magnetic resonance (NMR) spectroscopy analysis. The combined UPLC and NMR results allowed the identification of intermediates and side products.
Recommended Citation
Sulaitis, Katherine and Renner, Carine, "Ultra Performance Liquid Chromatography Analysis of Proteins and Synthetic Intermediates" (2016). Symposium on Undergraduate Research and Creative Expression (SOURCE). 541.
https://scholar.valpo.edu/cus/541
Biographical Information about Author(s)
Carine Renner is a junior biology and chemistry double major. She has a horse, and once was in Germany where she was filmed in a scene with George Clooney.
Katherine Sulaitis is a senior chemistry major, biology minor, and would like to solve murder cases with her powers of chemistry knowledge.
They became interested in this project because it is a unique experience provided for undergrads that allows them to operate an instrument new to the University (LC-MS).