User:Alexander Cvitan/Notebook/Experimental Biological Chemistry Lab/2013/09/03

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Objective

To calculate the molar absorptivity of adenosine and inosine by constructing a calibration curve. In addition, a Grubbs test will be performed on the class data in order to detect outliers, and remove them before data analysis. Procedure found here. <br.> Note that a q-test, and confidence intervals aren't going to be calculated like stated in the attached procedure. Instead a Grubbs test is going to be performed.

Procedure

Dilutions

Adenosine solution concentrations (M) Inosine solution concentrations (M)
3.00x10-5 4.80x10-5
2.50x10-5 4.00x10-5
2.00x10-5 3.20x10-5
1.50x10-5 2.40x10-5
1.00x10-5 1.60x10-5
0.50x10-5 0.80x10-5
0.25x10-5 0.4x10-5

These dilutions were made by making concentrated stock solutions of both Adenosine and Inosine.<br.>

Stock Solutions

  • .1091 grams of Adenosine was added to a volumetric flask and water was added to the 50 ml mark. This corresponds to a concentration of .0082M <br.>
  • .106 grams of Inosine was added to a volumetric flask and water was added to the 50 ml mark. This corresponds to a concentration of .0079M <br.>

Preparation of Dilutions

Screen shot 2013-09-03 at 3.00.44 PM.png<br.> All of the amounts listed above were measured using a micropipette. Water was then added to the amounts listed above in order to create a solution of 1ml. Note for the amount of concentrated solution added to each dilution, the micropipette is only accurate to the hundredths place. The water added to this solution was measured in a micropipette accurate to the tenths place.

Data

  • The graph below depicts the individual group absorbance spectra for adenosine.<br.>

Screen shot 2013-09-04 at 2.24.13 PM.png<br.>

  • Note Adenosine Absorbances had to be read twice because their was a smudge on the cuvette that gave odd absorbances for the first set of readings. We know it was something on the cuvette because the same samples were read the second time with a clean cuvette and it created the suspected calibration curve.<br.>
  • Peak at 259nm<br.>
  • Using the absorbances at the peak 259nm a calibration curve of our group data was created and depicted below

Adeno calibration Sept 4.png<br.>

  • Note we will be continuing this procedure on Sept 4, 2013.