Kinetics
Goals
- Test activity of HRP int he presence of AuNP by using a Uv-Vis Absorbance HRP assay.
Procedure
- We ran multiple UV-Vis spectras of the HRP with the gold nanoparticles to examine the impact that the NPs had on the kinetics of the reaction. We ran each run for 10 minutes and collected the various spectras. We maintained the same concentration in the cuvet of all the reagents except for the concentration of the gold nanoparticles. As a result, we were examining the effect that gold nanoparticles had on the rate of the reaction.
Data
- We ran 5 main tests and several of the tests multiple times to confirm the data. The multiple tests showed that the experiment was not really reproducible because we seemed to be getting different spectras every time.
| Trial run A
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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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| 1( standard) |
1.7 |
18 |
156.25 |
2.3 |
0 |
630
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| 1 |
1.7 |
18 |
156.25 |
2.3 |
20 |
610
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| 2 |
1.7 |
18 |
156.25 |
2.3 |
40 |
590
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| 3 |
1.7 |
18 |
156.25 |
2.3 |
60 |
570
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| 4 |
1.7 |
18 |
156.25 |
2.3 |
80 |
550
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| 5 |
1.7 |
18 |
156.25 |
2.3 |
100 |
530
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| Total volume added |
750 μL |
10 μL |
35 μL |
22 μL |
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The amount of HRP was changed from 22μL to 33μL because the 22μL in the cuvet did not give a linear spectra, just a horizontal line. More was needed in order to really push the reaction.
Excel spreadsheet of data.
File:HRPruns.xlsx
Conclusions
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Project name
