User:Alexander Cvitan/Notebook/Experimental Biological Chemistry Lab/2014/03/26

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Objective

  • UV-Vis and AA were run on room temperature lysozyme AuNP samples that we synthesized yesterday.
  • Based on today's Data: Due to the odd UV-Vis spectra observed for the AuNP's with added variable prepped yesterday we made new sets of AuNP's to ensure that a mistake wasn't made. Hopefully this data will agree with the data we previously reported or agree with the data we found today.

Procedure

Lysozyme-AuNPs

Creating Stock Solutions of Gold and Lysozyme

  • Stock solution were prepped using the info in the table below.
  • Each solution was measured using a volumetric flask and deionized water.


Synthesizing Lysozyme-AuNPs

  • As seen below, each solutions was created using the following calculations.
  • We made 66 tubes total.
  • Note: Instead of adding each amount into the tubes individually, we added the total volumes for 66 tubes into a beaker. We then measured 5 mL, which were transferred to glass test tubes.

Atomic Absorption

Creating the Gold Stock Solutions

  • Add 50 μL of HAuCl4·3H2O and 4950 μL of distilled water to a Falcon tube, for a final concentration of 10 μg/mL Au.
  • Add 100 μL of HAuCl4·3H2O and 4900 μL of distilled water to a Falcon tube, for a final concentration of 20 μg/mL Au.
  • Add 150 μL of HAuCl4·3H2O and 4850 μL of distilled water to a Falcon tube, for a final concentration of 30 μg/mL Au.
  • Add 200 μL of HAuCl4·3H2O and 4800 μL of distilled water to a Falcon tube, for a final concentration of 40 μg/mL Au.
  • Add 250 μL of HAuCl4·3H2O and 4750 μL of distilled water to a Falcon tube, for a final concentration of 50 μg/mL Au.


Atomic Absorption Samples

Solutions with the following Au:lysozyme ratio were run on the AA:

  • 30:1 lysozyme-AuNP with 0.03 M MgCl2, CaCl2, NaCl, KCl, MES, citric acid (0.0002316 M 2,2 bipyridine)
  • 30:1 lysozyme-AuNP with 0.06 M MgCl2, CaCl2, NaCl, KCl, MES, citric acid (0.0004544 M 2,2 bipyridine)
  • 30:1 lysozyme-AuNP with 0.09 M MgCl2, CaCl2, NaCl, KCl, MES, citric acid (0.0006772 M 2,2 bipyridine)

AA Graphs

  • This graph shows pretty constants amounts of gold present for all variables added. MES overall had the least amount of gold present in solution in comparison to other variables. Citric Acid was the only variable that a visual reduction in the amount of gold in solution as the concentration of variable increased.

UV-Vis Graphs

  • No 30:1 Lyso AuNP Control was Run because no samples were leftover from this batch of AuNP's. A single tube was made, but a mistake was made in its preparation causing fibers to form.

Image:Screen_Shot_2014-03-31_at_11.28.08_PM.png Image:Screen_Shot_2014-03-31_at_11.28.28_PM.png

  • This UV-Vis corroborates all of the data presented in the AA by showing a change in absorbance as concentrations of gold change.
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