User:Dhea Patel/Notebook/Hemoglobin Project/2013/07/17

From OpenWetWare

Jump to: navigation, search
Project name Main project page
Previous entry      Next entry

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

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

2. 26.0μL of the 47567μM solution of DAP in DMSO was added to 24.974mL of water to make a 50μM in a 25mL volumetric flask.

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


[STOCK] (μM) Initial Volume (mL) Vol. Water (mL) Final Volume (mL) [Final] (μM)
475670.10599.895100.00050.000
50.00015.00010.00025.00030.000
50.00013.50011.50025.00027.000
50.00012.00013.00025.00024.000
50.00010.50014.50025.00021.000
50.0009.00016.00025.00018.000
50.0007.50017.50025.00015.000
50.0006.00019.00025.00012.000
50.0004.50020.50025.0009.000
50.0003.00022.00025.0006.000
50.0001.50023.50025.0003.000
0.0000.00025.00025.0000.000

4. An absorbance spectrum was taken of each solution.

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

Concentration (μM) Absorbance
300.908
270.624
240.739
210.664
180.606
150.516
120.291
90.266
60.196
30.077
00

6. The slope of the best fit line is equal to the molar absorptivity (ε) of DAP in MeOH. ε = 29,400M-1cm-1.

7. Steps 1-6 were repeated in the afternoon. The following is the data obtained in the second trial. ε=20,800M-1cm-1.

Concentration (uM) Absorbance
300.647
270.555
240.517
210.429
180.37
150.291
120.23
90.179
60.14
30.061
00

8. The mixtures made during Trial 2 were run on the Shimadzu UV-2550 in the Beeghly 207. The following is the data obtained:

Concentration (uM) Absorbance
300.545
270.51
240.434
210.386
180.326
150.271
120.218
90.165
60.115
30.055
00


Personal tools