User:Elizabeth Ghias/Notebook/Experimental Biological Chemistry/2012/03/27

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Objective

To adjust the pHs of all the dye solutions (both first and second trial) so that they are all equal and to retake UV-Vis and fluorescence spectra

Description

1. The initial pHs of the solutions from both the first and second trials were measured.

First Trial Initial pHs

  • 70 molar ratio with dye pH = 8.12
  • 166 molar ratio with dye pH = 8.43
  • BSA/HCl with dye pH 8.30

Second Trial Initial pHs

  • 70 molar ratio with dye pH = 8.00
  • 166 molar ratio with dye pH = 8.13
  • BSA/HCl with dye pH = 7.98

2. Add 50 mM tris buffer (pH = 8.73) to raise the pHs of the samples to a pH around 8.30.

First Trial pH Changes

  • 70 molar ratio with dye: 10 drops of Tris were added to give a final pH of 8.30

Second Trial pH Changes

  • 70 molar ratio with dye: 6 drops of Tris added, final pH = 8.29
  • 166 molar ratio with dye: 3 drops of Tris added, final pH = 8.41
  • BSA/HCl with dye: 7 drops of Tris added, final pH = 8.28

3. The samples containing fibers (all dye samples) were centrifuged for 5 minutes at 13,200 rpg.

4. Fluorescence spectra were taken of the samples between 620 - 800 nm, with excitation at 600 nm.

5. Fluorescence spectra were also taken of the 70 molar ratio, 166 molar ratio, and the BSA/HCl reactions without dye.

6. UV-Vis spectra were taken of all dye and control solutions.


Data

Fluorescence Spectra of Solutions from the Second Trial

Image:27mar - fluorescence curve 2nd solution.jpg


Fluorescence Spectra of Solutions from the Initial Trial

Image:27mar - fluorescence curve initial solution.jpg


Fluorescence Spectrum of Control solutions

Image:27mar - fluorescence curve control solution.jpg


UV Spectra of Solutions from the Second Trial

Image:27mar - UV curve second solution.jpg

Image:27mar - UV curve second solution zoom.jpg


UV Spectra of Solutions from the Initial Trial

Image:27mar - UV curve initial solution.jpg

Image:27mar - UV curve initial solution zoom.jpg


UV Spectrum of Control Solutions

Image:27mar - UV curve control solution.jpg

Image:27mar - UV curve control solution zoom.jpg

Notes

  • When the pHs were initially measured, the values had changed from the week before. For example, the 166 molar ratio solution with dye from the second trial had a measured pH of 8.35 the previous week, but this week had a decreased pH of 8.13
  • The changes in pH values could be explained by contamination or a reaction that is occurring in the dye solutions.
  • The fluorescence spectra of the 70 molar ratio with dye now has a slightly higher intensity than that of the 166 molar ratio with dye. The BSA/HCl dye mixture now has the lowest intensity



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