Class: BE-210
Group: R5
Members: Vu Hong, Brett Riccio, Rebecca Lai, Chris Lindsey, Natalie Georgakis
Date: April 30, 1997
Full Text

Abstract:
Our objective in this experiment was to gain an understanding of the oxygenation of ferrohemoglobin. Hemoglobin is an important protein in the blood whose heme group enables it to carry oxygen to the cells in the body and take away their waste products. By studying how oxygenation relates to a peak weak wavelength through spectrophotometry, a method can be devised to detect the amount of oxygen in a patient's sample of blood. In specific, we investigated three things about oxyhemoglobin and deoxyhemoglobin: their peak wavelengths, the molar coefficients of each at their peak, and the relationship between their peak wavelengths and different concentrations of oxyhemoglobin. To achieve our goals, we analyzed three different samples of equine blood, performing three trials on each. To extract the hemoglobin , we used an IEC PR7000 Centrifuge to spin down the red blood cells and then lysed them with deionized water. Each sample was diluted so that its absorbance could be read on the Milton Roy Spectronic 20 Spectrophotometer for a range of wavelengths from 350-600nm. Then, we used the Beer Lambert Law to analyze our results. We found the peak wavelengths to be 410nm for oxyhemoglobin and 421nm for deoxyhemoglobin. In comparing these values to the literature values of 412nm and 420nm, our error was 0.485% and 2.093%, respectively. Secondly, we graphed our absorbance values versus concentration to find our molar coefficients to be 15.9*104 + 0.082 L/mol-cm and 15.1*104 + 0.151L/mol-cm. We had to compare these values with the coefficients at the literature peak wavelengths, 12.4*104 for oxyhemoglobin and 11.5*104 for deoxyhemoglibin, and found our error to be 28.47% and 31.30%, respectively. For the last objective, we found a linear relationship of an R squared value of 0.9426 between concentration of oxyhemoglobin and peak wavelength.