Honors Program


Date of this Version


Document Type



M. Gonzales, Evaluation of the Interactions Between Drugs and Binding Agents by Affinity Chromatography, Undergraduate Honors Thesis, University of Nebraska-Lincoln (2024).


Copyright Moira Gonzales 2024


Systems such as the human body or the natural environment involve interactions between multiple components in solution. Humic acid, a component of dissolved organic matter in soil, and human serum albumin (HSA), a transport protein in the body, can interact with drugs as binding agents. In this project, a method of high-performance affinity chromatography called ultrafast affinity extraction (UAE) was used to study the interactions between drugs and binding agents. The binding of ciprofloxacin and norfloxacin to Aldrich humic acid was studied, as well as the binding of repaglinide and warfarin to HSA. Solutions of the drug, the binding agent, and a mixture of the drug and binding agent were run at a range of flow rates for each of the four interactions being studied. This information was used to calculate the free fraction F0 of the drug in solution with the binding agent, as well as the rate dissociation constant kd and equilibrium constant Ka of the interaction.