User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/10/03

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Kinetics

Goals

  • Run UV-Vis of Absorbance vs. time of the HRP assay.
  • Abigail E. Miller 11:11, 7 October 2012 (EDT): that is what you did to accomplish your goal. yor goal is to test the activity of HRP in the presence of AuNPs 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.

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 ' ' ' ' ' '
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
1( standard) 1.7 18 156.25 2.3 0 630
1 1.7 18 156.25 2.3 20 610
2 1.7 18 156.25 2.3 40 590
3 1.7 18 156.25 2.3 60 570
4 1.7 18 156.25 2.3 80 550
5 1.7 18 156.25 2.3 100 530
Total volume added 750 μL 10 μL 35 μL 22 μL


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.


HRP with AuNPs


Excel spreadsheet of data.


File:HRPruns.xlsx

Conclusions

Graphs of the data should be found on Mary's notebook and analysis of data and rate reactions will be on in the future.