User:Moira M. Esson/Notebook/CHEM-571/2014/01/29

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Preparation of BSA-Magnetite solution)
(UV/vis)
Line 90: Line 90:
# After taking the spectrum, store the sample in a fresh eppy tube in an upright position.
# After taking the spectrum, store the sample in a fresh eppy tube in an upright position.
<br>
<br>
-
Figure 1. Corrected Absorbance Spectra of BSA-AuNP solutions (...ratios)
+
Figure 1. Corrected Absorbance Spectra of BSA-AuNP solutions (125:1-135:1 ratios)
<br>
<br>
 +
[[Image:CorrectedAbsorbancespectraof BSAAuproteinMTP.png]]
 +
<br>
 +
Figure 2. Corrected Absorbance Spectra of BSA-AuNP-Magnetite NP solutions (105:1-140:1)
 +
<br>
 +
[[Image:Magnetitegoldbsafiberprep1MTP.png]]
 +
<br>
 +
*This spectrum indicates a large concentration of iron in the solutions, yet lacks the characteristic gold peak. The fibers formed must be tested for magnetic properties.
 +
==Notes==
==Notes==
*A stir bar was placed in the reaction mixture. In the future, this will not be added.  
*A stir bar was placed in the reaction mixture. In the future, this will not be added.  

Revision as of 20:55, 3 February 2014

Project name Main project page
Previous entry      Next entry

Objectives

  1. Prepare a BSA-Magnetite solution with the purpose of synthesizing BSA fibers with magnetite nanoparticles incorporated in the fiber.
  2. Analyze protein-AuNP solutions prepared on 01/28/2014 using UV/vis.
  3. Analyze protein-AuNP-Magnetite solutions prepared on 01/28/2014 using UV/vis.


Preparation of BSA-Magnetite solution

  • The following protocol was adapted from the following reference[[1]]

General Protocol:

  1. Prepare a stock solution of BSA in a volumetric flask using deionized H2O.
  2. Prepare a 0.3M stock solution of FeSO4*7H2O in a 10mL volumetric flask using deionized H2O.
  3. Prepare a 1M stock solution of KOH in a 10mL volumetric flask using deionized H2O.
  4. Prepare a 0.8M stock solution of KNO3 in a 50mL volumetric flask using deionized H2O.
  5. Add the necessary amount of BSA for the desired ratio of BSA to Magnetite to a 10mL round bottom flask.
  6. Add the appropriate volume of FeSO4*7H2O to each test tube so that the solution has a concentration of 0.03M FeSO4*7H2O.
  7. Add the appropriate volume of KOH to each test tube so that the solution has a concentration of 0.1M KOH.
  8. Add the appropriate volume of KNO3 to each test tube so that the solution has a concentration of 0.4M KNO3.
  9. Add deionized H2O to the round bottom flask until the solution is 5mL. Seal the round bottom flask with a septum.
  10. Using a Schlenk line, de-gas the solution using nitrogen. De-gas the solution using nitrogen gas at room temperature for 5 minutes.
  11. Heat the reaction mixture to 90°C using a hot water bath for 4 hours. During this time, the reaction mixture should have a continuous flow of nitrogen gas.


  • Note: A previously prepared stock solution of 15mM BSA was used for the preparation.

Table 1. Preparation of BSA:MagnetiteNP solution

Ratio concentration BSA (M) volume of BSA added (mL) amount of FeSO4 added(mL) amount of KOH added (mL) amount of KNO3 added (mL) volume Water added (mL)
1300.0001153850.7354058980.50.5004459512.50.764148151


Table 2. Preparation of FeSO4*7H2O stock solution

Preparation of FeSO4 stock '
Molecular Weight (g/mol)278.01
mass(g)0.83403
volume stock (L)0.01
Concentration (M)0.3


Table 3. Preparation of KOH stock solution

Preparation of KOH stock '
Molecular Weight (g/mol)56.11
mass(g)0.5606
volume stock (L)0.01
Concentration (M)0.999108893


Table 4. Preparation of KNO3 stock solution

Preparation of KNO3 '
Molecular Weight (g/mol)101.1
mass(g)4.044
volume stock (L)0.05
Concentration (M)0.8


UV/vis

General Protocol:

  1. Place 1mL of each ratio in an 1.5mL eppendorf tube.
  2. Centrifuge at maximum speed for 30 minutes.
  3. Take a UV-Vis spectrum of each sample from 200nm to 800nm.
  4. After taking the spectrum, store the sample in a fresh eppy tube in an upright position.


Figure 1. Corrected Absorbance Spectra of BSA-AuNP solutions (125:1-135:1 ratios)
Image:CorrectedAbsorbancespectraof BSAAuproteinMTP.png
Figure 2. Corrected Absorbance Spectra of BSA-AuNP-Magnetite NP solutions (105:1-140:1)
Image:Magnetitegoldbsafiberprep1MTP.png

  • This spectrum indicates a large concentration of iron in the solutions, yet lacks the characteristic gold peak. The fibers formed must be tested for magnetic properties.

Notes

  • A stir bar was placed in the reaction mixture. In the future, this will not be added.
  • In comparison to the previously prepared solution, the solution remained a dark green color.
  • The solutions prepared on 01/28/2014 changed into a dark blue color. This is indicative of the preparation of magnetic magnetite particles.
  • A conglomeration of various fiber-esque precipitate formed in all of the prepared BSA:AuNP:MagnetiteNP solutions. Large particles also appear to be present. In comparison to the previously prepared protein fibers, these fibers appear much smaller (not as long or wide). The solutions will be filtered and these potential fibers will be collected for analysis.
  • The temperature of the hot water bath remained a 75-85°C. The temperature never reached 90°C.


Personal tools