Difference between revisions of "User:Eleni N. Kalivas/Notebook/CHEM-571/2013/09/18"

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(Redox Titration)
(Redox Titration)
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Redox Titration
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Redox titration of HRP % reduced data
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<br>
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''% Reduced'''
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| align="center" style="background:#f0f0f0;"|'''Potential'''
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|-
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| -2.73671656||0.0146
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|-
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| -0.472156173||-0.355
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|-
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| 0||-0.3519
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|-
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| 0.156439504||-0.3879
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|-
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| 56.60826937||-0.4045
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|-
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| 94.88970062||-0.4045
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|-
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| 96.52628268||-0.4781
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|-
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| 100||-0.5803
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|}
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<br>
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Revision as of 09:25, 1 October 2013

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Objective

  • The objective for today is to perform a redox titration on HRP in order to determine the standard redox potential of the protein.

Redox Titration

  • The protocol for this lab was found here.

Specific volume of samples used during redox titration.

Sample number Volume of HRP Sample(mL)
1 1
2 1.05
3 1.055
4 1.056
5 1.059
6 1.064
7 1.074
8 1.084

Absorbance Spectra of the redox titration of HRP FullyoxidizedreducedHRPzem.png

  • The maximum wavelength of fully oxidized HRP was 402.22nm, and the wavelength corresponding to the fully reduced HRP was 433.74nm. The sample with the addition of 64μL sodium dithionite was the fully oxidized sample, and the sample with the addition of 84uL sodium dithionite was the fully reduced sample. Since the reaction was not done in an entirely closed environment, oxygen from the atmosphere reacted with our sample which explains the large volumes added to our samples.


Potential of Redox titration of HRP PercentreducedHRPzem.png


Redox Titration of HRP

% Oxidized Potential
-2.633063185 -0.5122
0 -0.5803
3.260060544 -0.4781
30.97191805 -0.4045
82.70859316 0.0146
98.04493512 -0.3879
98.62977455 -0.355
100 -0.3519


Redox titration of HRP % reduced data

% Reduced Potential
-2.73671656 0.0146
-0.472156173 -0.355
0 -0.3519
0.156439504 -0.3879
56.60826937 -0.4045
94.88970062 -0.4045
96.52628268 -0.4781
100 -0.5803