User:Megan L. Channell/Notebook/Horseradish/2013/09/03

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Adenosine and Inosine UV-Vis

Objective

Finding the molar absorptivities of adenosine and inosine. This will help later on in studying adenosine deaminase (ADA) which converts adenosine to inosine.. The structural differences between the two molecules is that adenosine has a primary amine while inosine has a carboxy group.

Protocol

Stock Solutions

Two different stocks solutions were made, one adenosine and one inosine

  1. .0817g of adenosine was place in a 100mL volumetric flask and diluted with DI water
  2. .1261g of inosine was placed in a 100 mL volumetric flask and diluted with DI water
  3. The concentration of adenosine was determined to be 3.06mM and for inosine 4.70mM

Dilutions

A total of 14 dilutions were made

  1. Using the formula: M1V1=M2V2, the amount of stock solution needed was found.
  2. The sample of stock solution needed was pipetted into a 10mL volumetric flask and diluted with DI water
  3. This process was repeated 14 times

A sample calculation: Amount of Stock Solution.= [10 mL(dilution concentration)/(stock solution concentration)]

UV-Vis

  1. Absorption spectra was collected with DI water
  2. For each sample, 1mL of the sample was pipetted into a clean cuvette
  3. The sample was returned to the vial it was kept in and the cuvette was rinsed three times with DI water

Data

A UV-Vis was ran from 800-200nm. The absorbance and calibration curve were done through the data collected on the UV-Vis. The peak of Adenosine was at 259nm and the calibration curve was calculated off this wavelength.

Note

  • There was not enough time to run the UV-Vis on Inosine. Stock solutions and dilutions were stored for 9/4/2013.


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