User:Paul Rothenberg/Notebook/Pauls Notebook/2013/05/30

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Reaction of OPD and H2O2 in Water with Mb/Acetate Catalyst

  • Objectives:
  • 1. Continue analysis of reaction of OPD+H2O2 in Mb/Acetate Catalyst in aqueous solution
  • Procedure:
  • 1. Follow same procedure as 5/20 with Mb in Acetate buffer with H2O as the solvent
  • Data:

The protein did not fully dissolve in solution, similar to yesterday. However, the reaction still proceeded forwards. Some temperatures favored the reaction more than others, seen in the graphs below. The 25°C and 45°C reactions had particularly large absorbance spectrum at 420 nm, compared to the rest of the reactions. The 5°C and 15°C reactions did not have smooth curves; instead, had multiple peaks and relatively low absorbances, if any, at 420 nm. The 35°C reaction had a smooth curve but far less absorbance at 420 nm compared to 25°C and 45°C; with an absorbance of roughly 0.04 compared to 0.6 in the 45°C graph.



  • Not sure what happened at 15°C; the sequential graph is skewed compared to the rest.



  • At 3 minutes, the absorbance is higher between 500-550 nm, and then the later times decrease from this absorbance at this wavelength. There might be an intermediate product formed at that point.



Reaction of OPD and H2O2 in Water with Mb/Tris Catalyst

Continuing the analysis of this reaction with different solvents. Three of the four buffers have been tested, all with different results in the two different solvents.

  • Graphs:

  • Data:

The curves were overall smooth, as seen in the previous Tris reaction. The absorbance was not high compared to the other buffers used. The 35°C and 45°C reactions had the highest absorbances in this reaction, but even these absorbances were not as high as the Mb/Acetate run this morning.

In comparison to the reaction with Mb/Tris in MeOH, all of the reactions with H2O had a much higher absorbance than the reactions in MeOH.

Synthesis of Na12P2W18O62

Following the solution recrystallizing at 5°C overnight, we wanted to place it in the oven, as indicated in the protocol. Currently, it is still sitting in the fridge. Hopefully, we will fully dry it out and have the indicated 70g yield!

Update:

There is no functional vacuum oven in the building, so the solution in the crystals was removed and is being heated to recrystallize any more product and then filtered. The crystals will be placed in the oven at 80°C overnight and then vacuumed in a dessicator for a few hours to hopefully have the product. This is not an ideal situation, but we will see the results. Right now, there are copious amounts of light green crystals; hopefully the mass is close to 70 grams.


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