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)
47566.9501.00099.000100.000475.670
475.6701.0518.94910.00050.000
475.6700.9469.05410.00045.000
475.6700.8419.15910.00040.000
475.6700.7369.26410.00035.000
475.6700.6319.36910.00030.000
475.6700.5269.47410.00025.000
475.6700.4209.58010.00020.000
475.6700.3159.68510.00015.000
475.6700.2109.79010.00010.000
475.6700.1059.89510.0005.000
475.6700.0219.97910.0001.000
0.0000.00010.00010.0000.000

4. An absorbance spectrum was taken of each solution.

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

Concentration (uM) Absorbance
500.423
450.538
400.47
350.382
300.395
250.313
200.237
150.163
100.106
50.053
10.007
00

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.



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