User:Puja Mody/Notebook/AU Biomaterials 572/2014/02/11

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*UV-Vis
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Corrected Absorbance Spectra of BSA:Magnetite samples with differing concentrations of KNO<sub>3</sub>
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[[Image:MTP2112014.png]]
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*There is an increased absorbance at ~300nm for the sample with a higher concentration of KNO<sub>3</sub>. This lead to the conclusion that the absorbance at ~300nm corresponds with nitrate. The lack of an iron absorbance peak indicates that the Fe was successfully incorporated into the magnetite nanoparticles or that a  protein fiber was created.

Revision as of 15:15, 24 February 2014

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Day 5

  • Goals
    • UV-Vis the reaction mixtures prepared on 2/5
    • Prepare a new ratio of BSA- Magnetite solutions using both the oven method and the Schlenk line method with which to prepare them. This ratio will be the same as the hemoglobin- magnetite ratios prepared with the Schlenk line.
    • Analyze the magnetite nanoparticles which were dried Last week using FTIR
    • Dry the particulate collected from the reaction mixtures prepared on 2/5 so that they may be used for FTIR and XRD next week.

Procedure

  • We prepared the stock solutions of FeSO4*7H2O ,KNO3, and KOH in the same method and at the same concentrations as prepared on February 4
  • We made the following ten ratios for the oven method. the same procedure as used on 1/29

Table 1: Preparation of BSA and Magnetite solutions.

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)
105002.85714E-060.5685457660.50.5006245542.50.93082968
110002.72727E-060.5427027760.50.5006245542.50.95667267
115002.6087E-060.5191070030.50.5006245542.50.980268443
120000.00000250.4974775450.50.5006245542.51.001897901
125000.00000240.4775784430.50.5006245542.51.021797003
130002.30769E-060.4592100410.50.5006245542.51.040165405
135002.22222E-060.4422022620.50.5006245542.51.057173184
140002.14286E-060.4264093240.50.5006245542.51.072966122
145002.06897E-060.4117055540.50.5006245542.51.087669892
150000.0000020.3979820360.50.5006245542.51.10139341
  • The second method we used was the Schlenk Line method- the same as was utilized on 1/29. 2 samples were made at the following concentrations

Table 2: Hemoglobin stock solution

Preparation of hemoglobin Stock solution '
Molecular weight (g/mol)64500
mass hemglobin (g)0.0995
volume solution (L)0.1
Concentration (M)1.54264E-05

Table 3: Hemoglobin- Magnetite solution ratios with Schlenk Method

Ratio concentration Hemoglobin (M) volume of Hemoglobin added (mL) amount of FeSO4 added(mL) amount of KOH added (mL) amount of KNO3 added (mL) volume Water added (mL)
190001.57895E-060.5117693730.50.5006245542.50.987606073
210001.42857E-060.4630294330.50.5006245542.51.036346013

The Drying of samples- General Protocol

  1. Make sure the test tube/ round bottom flask the securely closed and mix vigorously the prepared samples until all precipitate is suspended in the solution.
  2. Pour the newly mixed solutions into a clean beaker (about a 10mL one)
  3. Place a magnet at the bottom/ side of the vial and wait for all the precipitate to to collect on the glass, out of the solution.
  4. One the solution has cleared, pour the solution out into a falcon tube while still holding the magnet at the bottom of the beaker.
  5. Place beaker with precipitate into fume hood and allow to air dry at least 24 hours.
    • To Run UV-Vis on the sample, do the following
  1. Centrifuge supernatant (collected solutions from above) for 30 minutes at 1500 rpm..
  2. Using ~1mL sample, analyze supernatant using UV/vis.

Data

For FTIR data and respective peaks please see Moira's Notebook



  • UV-Vis
Corrected Absorbance Spectra of BSA:Magnetite samples with differing concentrations of KNO3


Image:MTP2112014.png

  • There is an increased absorbance at ~300nm for the sample with a higher concentration of KNO3. This lead to the conclusion that the absorbance at ~300nm corresponds with nitrate. The lack of an iron absorbance peak indicates that the Fe was successfully incorporated into the magnetite nanoparticles or that a protein fiber was created.
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