User:Melissa Novy/Notebook/CHEM-571/2012/11/13

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   '''(0.00373 M)(V<sub>1</sub>)=(5 mL)(0.1×10<sup>-3</sup> M)'''
   '''(0.00373 M)(V<sub>1</sub>)=(5 mL)(0.1×10<sup>-3</sup> M)'''
   '''V<sub>1</sub> = 0.134 mL'''
   '''V<sub>1</sub> = 0.134 mL'''
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==UV-Vis Analysis of Au-HRP Solutions==
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* The Au-HRP solutions were made on [[User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/11/07|2012/11/07]] and the spectra of the solutions were obtained over 200 to 800 nm in quartz cuvettes.  Please refer to [[User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/11/13|Dhea Patel's entry]] for a photograph of the solutions.
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[[Image:UVvis_of_AUHRP_.png]]
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* Peaks at around 300 and 520 nm were visible.  The peak at 520 nm indicates the presence of GNPs in the 50, 100, and 150 mole ratio Au:HRP solutions.
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* The peak at around 300 nm was present in the spectra of the 250, 300, 350, 400, and 450 mole ratio Au:HRP solutions.  It is possible that this peak is due to the relatively higher concentration of HAuCl<sub>4</sub> in these solutions, based on [http://www.nanoscalereslett.com/content/pdf/1556-276X-5-124.pdf literature].  Note that the literature indicated that a peak at around 287 nm is present in the spectra of aqueous HAuCl<sub>4</sub>.
==Atomic Absorption of Au-Lysozyme and Au-HRP Solutions==
==Atomic Absorption of Au-Lysozyme and Au-HRP Solutions==

Revision as of 20:40, 29 November 2012

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Objectives

  • Analyze Au-HRP solutions with UV-vis spectroscopy.
  • Analyze solutions of Au-HRP and Au-lysozyme with AAS.
  • Conduct ADA activity assay on ADA fractions using adenosine.

ADA Activity Assay

  • 50 mL of 0.05 M sodium phosphate buffer was made with deionized water. The pH of the solution was adjusted to 7.4 using 3 drops of 12 M HCl.
  • 5 mL of 0.1 mM adenosine was made.
    • 100 mg of adenosine was dissolved in 1 mL of 0.05 M phosphate buffer, giving a solution of 0.00373 M adenosine.
    • 0.134 mL of 0.00373 M adenosine was diluted to a final volume of 5 mL with 0.05 M phosphate buffer, giving a solution of 0.1 mM adenosine.
  (0.001 g)/(268.02 g/mol) = 3.73×10-6 mol
  (3.73×10-6 mol)/0.001 L = 0.00373 M
  (0.00373 M)(V1)=(5 mL)(0.1×10-3 M)
  V1 = 0.134 mL

UV-Vis Analysis of Au-HRP Solutions

  • The Au-HRP solutions were made on 2012/11/07 and the spectra of the solutions were obtained over 200 to 800 nm in quartz cuvettes. Please refer to Dhea Patel's entry for a photograph of the solutions.

Image:UVvis_of_AUHRP_.png

  • Peaks at around 300 and 520 nm were visible. The peak at 520 nm indicates the presence of GNPs in the 50, 100, and 150 mole ratio Au:HRP solutions.
  • The peak at around 300 nm was present in the spectra of the 250, 300, 350, 400, and 450 mole ratio Au:HRP solutions. It is possible that this peak is due to the relatively higher concentration of HAuCl4 in these solutions, based on literature. Note that the literature indicated that a peak at around 287 nm is present in the spectra of aqueous HAuCl4.

Atomic Absorption of Au-Lysozyme and Au-HRP Solutions

Image:AAS_AuLys.JPG

Au-Lys Mole Ratio Au [ppm]
200.0057
301.9388
402.7586
503.806
604.5002
70-2.4599
80-2.4839
100-2.5856
120-2.6394
130-2.6454
  • Gold was detected in solutions at mole ratios of Au-lys from 20 to 60. The 60 Au-lys solution had the highest concentration of gold. This agrees with UV-vis data obtained on 2012/11/06. However, no peak or a slight peak was visible on the spectrum of the 20 Au-lys solution. This is corroborated by the fact that the AAS detected that the solution contained 0.0057 ppm Au.
  • It can be assumed that the fibers formed in the rest of the Au-lys solutions at mole ratios from 70 to 130 encapsulated the gold, such that there was no free gold or gold nanoparticles in solution. Theoretically, the gold concentration in these solutions was zero, but the AAS registered negative concentrations due to the fact that it was blanked with 1 N HCl, as opposed to deionized water.


Image:AAS_AuHRP.JPG

Mole Ratio of Au to HRP Au [ppm]
10-1.5443
501.8191
10011.7714
15013.1419
20023.6748
25024.4588
30027.876
35025.4761
40035.1173
45025.6198



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