User:Patrick Hampson/Notebook/chem471/2016/09/13

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BSA UV-Vis

The samples made in the previous lab period were flawed. Dr. Hartings remade the samples following these steps: (Link says unwritten).

Stock Solutions Used

  1. AuCl3
    1. mass = 34.62 mg
    2. 25.0 mL volumetric flask
    3. MW = 303.3 g/mol
    4. concentration = 4.57 mM
  2. Bovine Serum Albumin
    1. mass = 35.62 mg
    2. 10.0 mL volumetric flask
    3. MW = 66776 g/mol
    4. concentration = 53.3 mM
  3. Lysozyme
    1. 33.32 mg of lysozyme
    2. 10.0 mL water in volumetric flask
    3. MW = 14307 g/mol
    4. concentration = 232.9 μM
  4. Myoglobin
    1. 32.23 mg of myoglobin
    2. water to 10.0 mL in volumetric flask
    3. MW = 16951 g/mol
    4. concentration = 137 μM

Protocol

Detailed versions of the protocol for UV-Vis can be found in earlier notebooks. Samples were prepared with an identical concentration of gold. The pH of the solutions were fixed to be 4,5,6,7,8,9,10,11,12 using 1 mM HCL. The solutions were placed in a cuvette and run through the fluorimeter.

Data

160913 UV 0.125.PNG

This graph shows the absorbance of Fructose conc. 0.125 mM, pH 4-12 across 200-800 nm.

160913 UV 0.25.PNG

This graph shows the absorbance of Fructose conc. 0.25 mM, pH 4-12 across 200-800 nm.

160913 A530 0.125.PNG

This graph shows the absorbance at peak wavelength 530nm for Fructose concentration 0.125 mM (pH 4-12).

160913 A530 0.25.PNG

This graph shows the absorbance at peak wavelength 530nm for Fructose concentration 0.25 mM (pH 4-12).

Analysis

Similar to the UV-Vis trial the group previously conducted, the samples of pH 11 and 12 (for both concentrations of fructose) had significantly lower absorbance than that of the samples with pHs 4-10. Fructose 0.25 with pH 11 and 12 had a very faint pink coloration to them. Samples with pH 4-10 all had similar absorbance across the spectrum. Max absorbance at 530 nm was also similar.



Notes

160913 0.125 AU NP

0.125 mM Fructose, 0.25 mM Au samples of pH 4-12.

160913 0.25 NP

0.25 mM Fructose, 0.25 mM Au samples of pH 4-12.