User:Moira M. Esson/Notebook/CHEM-571/2014/02/05

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(FTIR and XRD Sample Preparation)
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==UV/vis==
==UV/vis==
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*All samples prepared on [[User:Moira M. Esson/Notebook/CHEM-571/2014/02/04|02/04/2014]] were analyzed using UV/vis spectroscopy.
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*The samples were not centrifuged, as the nanoparticles/fibers were removed from the solution in preparation for FTIR and XRD analysis prior to UV/vis analysis.
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<br>
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'''Figure 1.''' Corrected Absorbance Spectra of Hemoglobin:Magnetite nanoparticles/fibers
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<br>
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[[Image:Corrected Absorbance Spectra of Heme-Magnetite nanoparticles .png]]
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<br>
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*Strong absorbance peak around 302nm corresponds to the excess nitrate present from the magnetite synthesis. The weak absorbance peaks
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'''Figure 2.''' Corrected Absorbance Spectra of BSA:Magnetite nanoparticles/fibers
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<br>
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[[Image:Corrected Absorbance Spectra of BSA-Magnetite nanoparticles .png]]
==Observations/Notes==
==Observations/Notes==

Revision as of 21:15, 10 February 2014

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Objective

  1. Prepare all magnetite samples that were synthesized on 01/28/2014 and 01/29/2014 for FTIR and XRD analysis.
  2. Analyze magnetite nanoparticle/protein fiber samples prepared on 02/04/2014 using UV/vis spectroscopy.
  3. Synthesize two different ratios of BSA:Magnetite nanoparticles/fibers


BSA:Magnetite nanoparticle preparation

  • The general protocol described on 01/29/2014 was followed.
  • Two solutions were prepared. Both solutions were prepared as a 13000:1 BSA:Magnetite.
    • The difference between the two solutions is the concentration of KNO3 that was added to the solution.
    • Reaction mixture 1 contains the concentration of KNO3 that follows the protocol adaptation from the following reference [[1]].
    • Reaction mixture 2 contains the concentration of KNO3 described on 02/04/2014.


Table 1. Preparation of both reaction mixtures

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)
130002.30769E-060.4592100410.50.5006245542.51.040165405


Table 2. Preparation of BSA stock solution

Preparation of BSA stock '
Molecular weight (g/mol)66463
mass (g)0.0167
volume stock (L)0.01
Concentration (M)2.51268E-05


Table 3. Preparation of KOH stock solution

Preparation of KOH stock '
Molecular Weight (g/mol)56.11
mass(g)0.5604
volume stock (L)0.01
Concentration (M)0.998752451


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 KNO3 stock solution for Reaction Mixture 1

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


Table 6. Preparation of KNO3 stock solution for Reaction Mixture 2

Preparation of KNO3 '
Molecular Weight (g/mol)101.1
mass(g)0.5604
volume stock (L)0.01
Concentration (M)0.5


FTIR and XRD Sample Preparation

  • All samples that were centrifuged on 02/04/2014 were prepared for FTIR and XRD analysis.
  • Two samples prepared using the Schlenk line method on 02/04/2014 and [[01/29/2014

General protocol for centrifuged samples:

  1. Transfer all supernatant from falcon tube into a clean, labeled falcon tube using a disposable Pasteur pipette. Store the supernatant for future AA analysis.
  2. Using a spatula, scrap precipitate and place into a clean 10mL beaker.
  3. To ensure all precipitate (this precipitate is either magnetite nanoparticles or protein fibers with magnetite nanoparticles) add no more than 2mL deionized H2O to falcon tube and vortex falcon tube for 1 minute.
  4. Pour contents of falcon tube into beaker.
  5. Label and place beaker in fume hood to completely dry.


General protocol for samples prepared using Schlenk line method:

  1. Pour reaction mixture into a 25mL beaker. (Stir the reaction mixture prior to pouring to ensure all nanoparticles are part of a colloid suspension)
  2. Using a magnet, collect all nanoparticles at the bottom of the beaker. Hold the magnet on the bottom of the beaker for ~3 minutes or until the solution is completely transparent/colorless.
  3. Gently pour the supernatant into a clean, labeled falcon tube. While pouring, ensure that the magnet is still held at the bottom of the beaker so as no magnetic nanoparticles are poured into the falcon tube.
  4. Place beaker in fume hood and allow to completely dry.

UV/vis

  • All samples prepared on 02/04/2014 were analyzed using UV/vis spectroscopy.
  • The samples were not centrifuged, as the nanoparticles/fibers were removed from the solution in preparation for FTIR and XRD analysis prior to UV/vis analysis.


Figure 1. Corrected Absorbance Spectra of Hemoglobin:Magnetite nanoparticles/fibers
Image:Corrected Absorbance Spectra of Heme-Magnetite nanoparticles .png

  • Strong absorbance peak around 302nm corresponds to the excess nitrate present from the magnetite synthesis. The weak absorbance peaks

Figure 2. Corrected Absorbance Spectra of BSA:Magnetite nanoparticles/fibers
Image:Corrected Absorbance Spectra of BSA-Magnetite nanoparticles .png

Observations/Notes

  • The Hemoglobin:magnetite colloid suspension/fibers prepared on 02/04/2014 formed larger particles than the previously prepared solution using the Schlenk line. The resulting reaction mixture was clear other than the formed particles at the bottom of the round bottom flask. Using a magnet, the particles from the reaction mixture prepared on 02/04/2014 appeared to be more magnetic (i.e. were more easily moved when an external magnetic field was applied) than the particles prepared on 01/29/2014. This may be due to the different ratio of protein to FeSO4 used. As such, two new BSA:magnetite solutions were prepared in order to compare.
  • Due to the absence of an iron lamp for AA, all reaction mixture solutions were saved and stored (after removal of precipitate for XRD and FTIR analysis) for future AA analysis.



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