User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/03

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(→Kinetics of AuNP (Citrate))
Current revision (16:49, 11 December 2012) (view source)
(→Kinetics of AuNP (Citrate))
 
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* Now that the optimal amount of HRP was found, the reaction of AAP and hydrogen peroxide with HRP was tested if this was affected by the addition of Au/Citrate. The first run is HRP alone with the reaction followed by the addition of Au/Citrate.
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* Now that the optimal amount of HRP was found, the reaction of AAP and hydrogen peroxide with HRP was tested if this was affected by the addition of Au/Citrate. The first run is HRP alone with the reaction followed by the addition of Au/Citrate as shown below.
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* Au/Citrate was made from 20 mL of 1 mM HAuCl<sub>4</sub> with 20 mL of 1% citrate. The Au/Citrate was added in multiple of 20. Each 20 μL is equivalent to 2%.
[[Image:Aucit.png]]
[[Image:Aucit.png]]
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* The amount and concentration of reactants are shown below.
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Trials'''
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| align="center" style="background:#f0f0f0;"|''''''
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| align="center" style="background:#f0f0f0;"|''''''
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| align="center" style="background:#f0f0f0;"|''''''
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| align="center" style="background:#f0f0f0;"|''''''
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| align="center" style="background:#f0f0f0;"|''''''
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| align="center" style="background:#f0f0f0;"|''''''
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|-
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| Cuvet||hydrogen peroxide concentration (mM)||Iodophenol in DMSO concentration (μM)||AAP Concentration (μM)||HRP concentration (μM)||μL of gold np added||μL of water added
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|-
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| 1( standard)||1.7||18||156.25||3.45||0||630
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|-
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| 1||1.7||18||156.25||3.45||20||610
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|-
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| 2||1.7||18||156.25||3.45||40||590
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|-
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| 3||1.7||18||156.25||3.45||60||570
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|-
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| 4||1.7||18||156.25||3.45||80||550
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|-
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| 5||1.7||18||156.25||3.45||100||530
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|-
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| Total volume added||750 μL||10 μL||35 μL||33 μL||||
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|}
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Kinetics of AuNP (Citrate)

  • As mentioned in the previous entry of Oct. 2, there was difficulty in finding the appropriate ratio between AAP and HRP.
  • Using the Kinetics program, trial and error was done to find a linear trend for the absorbance of HRP. The calculated volume was 22 μL. The optimal volume for HRP was found at 33 μL. This can be seen on the graph below.

Image:HRP.png


  • Now that the optimal amount of HRP was found, the reaction of AAP and hydrogen peroxide with HRP was tested if this was affected by the addition of Au/Citrate. The first run is HRP alone with the reaction followed by the addition of Au/Citrate as shown below.
  • Au/Citrate was made from 20 mL of 1 mM HAuCl4 with 20 mL of 1% citrate. The Au/Citrate was added in multiple of 20. Each 20 μL is equivalent to 2%.


Image:Aucit.png


  • The amount and concentration of reactants are shown below.
Trials ' ' ' ' ' '
Cuvethydrogen peroxide concentration (mM)Iodophenol in DMSO concentration (μM)AAP Concentration (μM)HRP concentration (μM)μL of gold np addedμL of water added
1( standard)1.718156.253.450630
11.718156.253.4520610
21.718156.253.4540590
31.718156.253.4560570
41.718156.253.4580550
51.718156.253.45100530
Total volume added750 μL10 μL35 μL33 μL



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