A half-semester, undergraduate research project was designed for an upper level course on separation methods. The main objective of the project was to determine the parameters (i.e., mobile phase composition and flow rate) for optimal separation (i.e., high resolution and low analysis time) of a three-component mixture by reverse-phase HPLC using published solvents as the mobile phase and C18 as the stationary phase. Students were expected to work independently for a minimum of three consecutive hours per week over eight weeks. An outline of tasks for each week was provided as a means to help students gauge their progress. Week one, perform a literature search for an HPLC chromatogram obtained using C18. Week two, construct ChemDraw and Spartan models of each analyte, calculate analyte hydrophobicity (i.e., Log P), and hypothesize elution order. Week three, prepare each solvent of the mobile phase separately, and prepare HPLC samples for each analyte. Week four, analyze the sample of each analyte using the published HPLC parameters. Week five, prepare a three-component mixture of the analytes, and analyze the mixture using different mobile phase compositions. Week six, analyze the mixture using different mobile phase flow rates. Week seven, perform quantitative analysis of each analyte by preparing a standard curve of a set of standards prepared by serial dilution. Week eight, prepare a mock journal article and a mock graduate research update. By the end of the project, students have gained a better understanding on the concepts of chromatography, optimizing resolution, and HPLC, developed lab experience in HPLC analysis and method development, and fostered communication skills in writing and speaking.

Design of a half-semester, undergraduate research project: Optimization of the separation of a three component mixture by reverse-phase HPLC using C18

ACS Spring Meeting 2021

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.