User:Dhea Patel/Notebook/Hemoglobin Project/2013/07/16

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Finding the Molar Absorptivity of 2,3-Diaminophenazine (DAP) in Water

1. 10.0mg of 2,3-Diaminophenazine (210.23 g/mol) was added to 1.0mL of DMSO to make a 47567μM solution.

2. 1.0 mL of the 47567μM solution of DAP in DMSO was added to 99.0mL of water to make a 475.67μM in a 100mL volumetric flask.

3. The following table outlines the volumes of DAP stock used and the final concentration of DAP solutions created.

[STOCK] (uM) Initial Volume (mL) Vol. Water (mL) Final Volume (mL) [Final] (uM)
47566.950 1.000 99.000 100.000 475.670
475.670 1.051 8.949 10.000 50.000
475.670 0.946 9.054 10.000 45.000
475.670 0.841 9.159 10.000 40.000
475.670 0.736 9.264 10.000 35.000
475.670 0.631 9.369 10.000 30.000
475.670 0.526 9.474 10.000 25.000
475.670 0.420 9.580 10.000 20.000
475.670 0.315 9.685 10.000 15.000
475.670 0.210 9.790 10.000 10.000
475.670 0.105 9.895 10.000 5.000
475.670 0.021 9.979 10.000 1.000
0.000 0.000 10.000 10.000 0.000

4. An absorbance spectrum was taken of each solution.

5. The absorbance at 420.1nm was plotted vs. the solution concentrations.

Concentration (uM) Absorbance
50 0.423
45 0.538
40 0.47
35 0.382
30 0.395
25 0.313
20 0.237
15 0.163
10 0.106
5 0.053
1 0.007
0 0
07162013 Molar Absorptivity of DAP in H2O Trial 2 GRAPH.png

6. The slope of the best fit line is equal to the molar absorptivity (ε) of DAP in water. ε = 11,000M-1cm-1.

  • [Fu et. al.] calculated ε of DAP in water to be 13,000M-1cm-1.