User:Puja Mody/Notebook/AU Biomaterials 572/2014/01/28

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==Observations==
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*The addition of the FeSO<sub>4</sub>*7H<sub>2</sub>O  solution to the gold caused an immediate formation of the gold nanoparticles through the reduction of the free gold by the  FeSO<sub>4</sub>*7H<sub>2</sub>O. This reaction was depicted by an immediate change in the color of the sample solutions from a yellowish hue (characteristic of the gold solution) to a deep/ dark purple. 

Revision as of 22:21, 3 February 2014

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Day 1

  • Goals:
  1. Prepare 3 Bovine Serum Albumin (BSA) and gold nanoparticle protein fibers as a standard for comparison against the solutions containing BSA, Gold, and magnetite.
  2. Determine the ideal concentration necessary to prepare BSA protein fibers with gold and magnetite nanoparticles by preparing 10 solutions of varying concentration ratios.

Procedure

  • Preparation protocol for AuNPs in BSA solution.
  1. Prepare separate stock solution of each BSA and HAuCl4 in volumetric flasks deionized water.
  2. Add the necessary volumes of gold and BSA to achieve the desired concentration ratios (BSA:AU) into 10 test tubes. The total volume of the samples should not exceed 5mL.
  3. Add the necessary amount of deionized H2O into the test tubes in order to achieve a final volume in each test tube of 5mL
  4. Cap the test tubes with an aluminum cap and heat in an oven for 3 hours at 80°C.
  5. Store the remaining solutions in falcon tubes.


  • Preparation of Magnetite-AuNP-BSA Solutions
    • These solutions were prepared following the same method as the BSA-AuNP solutions.
    • The following concentrations of FeSO4*7H2O, KOH, and KNO3 in each prepared solution were adapted from the following reference[[1]]


Protocol:

  1. Prepare the following two stock solutions in a 10mL volumetric flask using deionized H2O. Prepare KNO3 in a 50mL volumetric flask.

- 0.3M solution of FeSO4*7H2O - 1M solution of KOH i - 0.8M solution of KNO3.

  1. Add the appropriate volume of BSA and gold in a 10mL test tube to obtain the desired ratio of BSA:Au. Adjust levels so that the total volume of the solution does not exceed 5mL at this time.
  2. Add the appropriate volume of FeSO4*7H2O to each test tube so that the solution has a concentration of 0.03M FeSO4*7H2O.
  3. Add the appropriate volume of KOH to each test tube so that the solution has a concentration of 0.1M KOH.
  4. Add the appropriate volume of KNO3 to each test tube so that the solution has a concentration of 0.4M KNO3.
  5. Add deionized water to all of the test tube samples in order to bring up all the solutions to 5mL. Cap the test tubes with aluminum caps and heat them in the oven for 3 hours at 80°C.
  6. Store any remaining stock solutions in falcon tubes.

Note:Use caution when removing the test tube from oven. The test tubes will be hot.


Calculations

  • Preparation of BSA and Gold.

Table 1. Preparation of BSA Stock solution

Preparation of BSA stock '
Molecular weight (g/mol)66463
mass (g)0.0164
volume stock (L)0.01
Concentration (M)2.46754E-05


Table 2. Preparation of Gold Stock solution

Preparation gold stock '
Molecular weight Au (g/mol)393.83
mass Au(g)0.0326
volume stock(L)0.025
Concentration (M)0.003311073


Table 3. Preparation of BSA-AuNP solutions- We chose these three ratios based on the knowledge of the fiber producing region from previous experiement.s

Ratio Amount of gold added (mL) Volume BSA added (mL) Volume water added (mL)
1250.3775210890.4052621954.217216716
1300.3775210890.3896751884.232803723
1350.3775210890.3752427734.247236138



  • Preparation of Magnetite, AuNPs, and BSA


Table 4. 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 5. Preparation of the 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 6. 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


Table 7. Preparation of the Magnetite nanoparticle:AuNP:BSA solutions. Because we did not know where to expect fibers to form. We chose a wide range of ratios to test.

Ratio Amount of gold added(mL) concentration BSA (M) volume BSA (mL) amount of FeSO4 added(mL) amount of KOH added (mL) amount of KNO3 added (mL) volume Water added (mL)
1050.3775210892.38095E-060.4824549940.50.5004459512.50.639577966
1100.3775210892.27273E-060.4605252220.50.5004459512.50.661507739
1150.3775210892.17391E-060.4405023860.50.5004459512.50.681530574
1200.3775210892.08333E-060.422148120.50.5004459512.50.69988484
1250.3775210890.0000020.4052621950.50.5004459512.50.716770765
1300.3775210891.92308E-060.3896751880.50.5004459512.50.732357773
1350.3775210891.85185E-060.3752427730.50.5004459512.50.746790187
1400.3775210891.78571E-060.3618412460.50.5004459512.50.760191715
1450.3775210891.72414E-060.3493639610.50.5004459512.50.772668999
1500.3775210891.66667E-060.3377184960.50.5004459512.50.784314464


Observations

  • The addition of the FeSO4*7H2O solution to the gold caused an immediate formation of the gold nanoparticles through the reduction of the free gold by the FeSO4*7H2O. This reaction was depicted by an immediate change in the color of the sample solutions from a yellowish hue (characteristic of the gold solution) to a deep/ dark purple.



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