User:Hannah Harvey/Notebook/CHEM-471 Fall 2016/2016/09/13

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Objectives

  1. Run fluorescence data for lysozyme at pH=5
  2. Create stock solutions for use on 09.14.2016

Description

Fluorescence

  1. Solutions made using measurements below
  2. Fluorescence data run every 5 minutes for 3 hours

Solutions for 09/14/2016
Solutions were added to glass test tubes, covered with foil, and placed in an oven set to 80°C for 4 hours.

Data

Fluorescence

  1. Solution Measurements
    1. Lysosome Stock Solution
      1. 33.32 mg of lysozyme with 10.0 mL of water
      2. MW = 14307 g/mol
      3. concentration = 233 μM
    2. Gold Stock Solution
      1. 34.62 mg of AuCl3 with 25.0 mL of water
      2. concentration = 4,57 mM
    3. Lysozyme: 0.25mM Au, 12.5μM Lysozyme, 3mL total; pH 5
      1. 164 μL AuCl3
      2. 30 μL 1M HCl
      3. 161 μL Lysozyme
      4. 2645 μL Water
  2. Fluorescence Summary Graphs
091416 Emission v time lysozyme pH5.png

Figure 1 Maximum emission data of tryptophan in a solution of pH 5.

091416 Integrated v time Lysozyme pH5.png

Figure 2 Integrated fluorescence data of tryptophan in a solution of pH 5.

091316 Full Spectra pH5.png

Figure 3 Full emission spectra for important time points: t=0, 5, 35, 90, 180 min.

Figure 3 Explanation

  1. At t=0 min, we see a max emission far to the left of the other emission spectra.
  2. At t=5 min, this maximum has shifted up and to the right.
  3. Until t=35 min, the maximum for each spectra decreases until the shown maximum.
  4. After t=35 min, the maximum begins increasing again.
  5. At t=90 min, the maximum continues to increase, but the "shoulder" seen at around 475 nm begins going away.
  6. t=180 shows the final full spectra obtained.

UV-vis solutions for 09.14.2016
091216 UV-vis sol fruc 125.png
Table 1 UV-vis solutions made for [Fructose]=.125mM

091216 UV-vis sol fruc 25.png
Table 2 UV-vis solutions made for [Fructose]=.25mM

Notes

At t=0, the cuvette had been in the machine briefly for about 1-2 minutes before the machine was started, which could account for some error in the data.
While running the fluorescence data, the machine briefly shut off at the 100 minute mark. The fluorescence data shows t=95, t=103, t=105 instead.

After taking the UV-vis samples out of the 80°C oven, pH 11 for both Fructose concentrations showed a precipitate form. pH 12 did not show this precipitate.