User:James C. Schwabacher/Notebook/CHEM-571/2013/09/17

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Objective

  • Find the endpoint for 100% oxidation of Horseradish Peroxidase solution with potassium ferricyanide
  • Find the endpoint for 100% reduction of Horseradish Peroxidase solution with sodium dithionite

Procedure

  1. Procedure followed according to Prof. Hartings. During the procedure it was decided to add the oxidation and reduction reagents by increments of 2 uL instead of 1uL.

Data

  • Image:CHEM571_09.17.13_potassiumHRP_UVVIS.png
  • Image:CHEM571_09.17.13_sodiumdiHRP_UVVIS.png
  • Image:CHEM571_09.17.13_potassiumHRP.png
  • Image:CHEM571_09.17.13_sodiumdiHRP.png

Notes

As a whole, the class failed to see evidence of UV-vise shifting due to oxidation. It is believed that the HRP solution was already partially oxidized prior to oxidation.

Furthermore, there was similar difficulty when collecting data for the reduction reaction. More research is needed into the errors that occurred today.


Update: Prof. Hartings found that the iron in the protein was likely in the 3+ oxidation state, thus requiring 2000 equivalents of sodium dithionite to be added for full reduction to occur. His findings can be found here.



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