User:Alexander Cvitan/Notebook/Experimental Biological Chemistry Lab/2013/10/08

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Objective

  • To complete the procedure outline by Dr.Hartings HERE.
  • We hope to outline the kinetics of the conversion of a solution of Adenosine to Inosine using ADA.

Procedure

  • We prepared a 3ml sample solution of 40uM Adenosine, using 3ul of 40mM Adenosine and 2997 ul buffer. <br.>
  • The 40 mM sample was prepared using .1069 grams of Adenosine in 10 ml of buffer.<br.>
  • 30 ul of ADA was added after the fifth run was recorded in the data. Data was recorded every 15 sec for 10 min.<br.>
  • Dilutions were made with 50mM Phosphate buffer pH 7.4
  • ADA Stock Solution 0.11 units/mL ADA

Data

Spectra of ADA Conversion of Adenosine to Inosine

  • This graph represents the conversion of Adenosine to Inosine using ADA. Shown in the graph, there is a shift in the spectra which were recorded every 15 sec for 10 min.
  • Overall it was observed that the adenosine peak shrunk overtime, and everything appeared to shift to the right.

Oct 8 adenosine inosine kinetics.png

  • At peak 250 nm; the molar absorptivity of adenosine and inosine are 10224 (1/M*cm) and 11007 (1/M*cm) respectively.
  • At Peak 260 nm; the molar absorptivity of adenosine and inosine are 14025 (1/M*cm) and 6630 (1/M*cm) respectively.

Concentration vs. Time for ADA Conversion of Adenosine to Inosine

    • The graph below represents the change in concentration from adenosine to inosine over time using ADA. In order to calculate this from the spectra above, molar absorptivities from a previous day were utilized. As seen in the graph it isn't perfect because the concentration of inosine at time zero should ideally be zero.

Oct 8 Con vs. time ADA.png