Difference between revisions of "User:Moira M. Esson/Notebook/CHEM-571/2013/11/06"

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==Entry title==
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==Objectives==
 
#Continue biomineralization. Prepare protein-AuNPs for Hemoglobin and Lysozyme.
 
#Continue biomineralization. Prepare protein-AuNPs for Hemoglobin and Lysozyme.
 
#Analyze protein-AuNP solutions prepared on [[User:Moira M. Esson/Notebook/CHEM-571/2013/10/30|10/30/2013]] using UV/vis spectroscopy.
 
#Analyze protein-AuNP solutions prepared on [[User:Moira M. Esson/Notebook/CHEM-571/2013/10/30|10/30/2013]] using UV/vis spectroscopy.
 
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==UV/vis Analysis==
 
==UV/vis Analysis==
 
'''General Protocol''':
 
'''General Protocol''':

Revision as of 11:50, 19 November 2013

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Objectives

  1. Continue biomineralization. Prepare protein-AuNPs for Hemoglobin and Lysozyme.
  2. Analyze protein-AuNP solutions prepared on 10/30/2013 using UV/vis spectroscopy.


UV/vis Analysis

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 Lysozyme-AuNP solutions

Corrected Absorbance Spectra of Lysozyme-AuNP solutions.png

  • Only the 30:1 ratio exhibited an absorbance peak at 526nm. This was expected as this was the only colored solution or only solution that did not form fibers shown on 11/05/2013


Figure 2. Corrected Absorbance Spectra of Myoglobin Au-NP solutions
Corrected Absorbance spectra of Myoglobin Au-NP solution .png

  • 15:1 ratio is the only exhibited an aborbance peak at 732nm. This may indicate the optimal concentration for the preparation of fibers. Although this solution produced fibers 11/05/2013 this solution is still easily monitored using UV/vis spectroscopy.

Protein-AuNP solution preparation

  • The general protocol for the synthesis of Hemoglobin-AuNP and Lysozyme-AuNP follows the general protocol for the preparation of protein wrapped Au-NPs described on 08/28/2013.
  • 5mL of each solution was prepared.
  • All solutions were prepared using deionized H2O
  • Tri-hydrate gold was used for the preparation of the gold standard solution.
  • All stock solutions were prepared in a 10mL volumetric flask.


Table 1. Preparation of Hemoglobin Stock solution

Molecular weight (g/mol) 64500
mass myoglobin (g) 0.0106
volume solution (L) 0.01
Concentration (M) 1.64341E-05


Table 2. Preparation of Gold stock solution

Molecular weight Au (g/mol) 393.83
mass Au(g) 0.032
volume stock(L) 0.025
Concentration (M) 0.003250133


Table 3. Preparation of Lysozyme stock solution

Molecular weight (g/mol) 14307
mass lysozyme (g) 0.0717
volume solution (L) 0.05
Concentration (M) 0.00010028


Table 4. Preparation of Hemoglobin-AuNP solutions

Ratio Amount of gold added concentration hemoglobin (M) volume Hemoglobin (mL) Volume Water (mL)
30 0.384599609 8.33333E-06 2.535377358 2.080023032
50 0.384599609 0.000005 1.521226415 3.094173976
70 0.384599609 3.57143E-06 1.086590296 3.528810094
90 0.384599609 2.77778E-06 0.845125786 3.770274604
110 0.384599609 2.27273E-06 0.691466552 3.923933838
130 0.384599609 1.92308E-06 0.585087083 4.030313308
150 0.384599609 1.66667E-06 0.507075472 4.108324919
170 0.384599609 1.47059E-06 0.447419534 4.167980857
190 0.384599609 1.31579E-06 0.400322741 4.21507765
210 0.384599609 1.19048E-06 0.362196765 4.253203625


Table 5. Preparation of Lysozyme Au-NP solutions

Ratio Amount gold added (mL) Concentration Lysozyme (M) Volume lysozyme (mL) Volume water (mL)
5 0.384599609 0.00005 2.493019545 2.122380845
10 0.384599609 0.000025 1.246509773 3.368890618
15 0.384599609 1.66667E-05 0.831006515 3.784393876
20 0.384599609 0.0000125 0.623254886 3.992145504
25 0.384599609 0.00001 0.498603909 4.116796482
30 0.384599609 8.33333E-06 0.415503258 4.199897133
35 0.384599609 7.14286E-06 0.356145649 4.259254741
40 0.384599609 0.00000625 0.311627443 4.303772947
45 0.384599609 5.55556E-06 0.277002172 4.338398219
50 0.384599609 0.000005 0.249301955 4.366098436
  • New ratios of lysozyme were prepared in order to determine the optimal concentration ratios for the formation of fibers.