Binding of Oxygen to Human Hemoglobin Within the Erythrocyte Using ICAM Spectrophotometry

Authors

Kyle K. Hill, University of Nebraska-LincolnFollow

Date of this Version

Spring 4-25-2016

Citation

Hill, K. (2016). Binding of Oxygen to Human Hemoglobin within the Erythrocyte using ICAM Spectrophotometry (Master's Thesis). Available in Digital Commons, University of Nebraska, Lincoln, Nebraska.

Comments

A THESIS Presented to the Faculty of The Graduate College at the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Master of Science, Major: Chemistry, Under the Supervision of Professor Lawrence J. Parkhurst. Lincoln, Nebraska: April, 2016

Copyright © 2016 Kyle Kelly Hill

Abstract

Many of the spectrophotometric techniques used to determine the properties of intracellular human hemoglobin cannot be utilized due to the turbidity of erythrocyte suspensions. An Integrating Cavity Absorption Meter, or ICAM, allows for absorption measurements of strongly scattering samples in the visible-light region of the spectrum. The spectrum of oxygenated hemoglobin within erythrocytes is significantly different from the absorption spectrum of oxygenated hemoglobin in solution. Studies of the oxygen binding to hemoglobin in erythrocytes allowed the four sequential binding constants (Adair constants) to be determined and compared with those of hemoglobin in solution. The Adair constants for hemoglobin in solution were found to be markedly different from those of hemoglobin in the erythrocyte.

Advisor: Lawrence J. Parkhurst