User:Moira M. Esson/Notebook/CHEM-571/2013/09/03: Difference between revisions

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# Prepare inosine solutions of various concentrations to determine the molar absorptivity of [http://en.wikipedia.org/wiki/Inosine inosine].
# Prepare inosine solutions of various concentrations to determine the molar absorptivity of [http://en.wikipedia.org/wiki/Inosine inosine].
# Prepare calibration curves for adenosine and inosine.
# Prepare calibration curves for adenosine and inosine.
# Analyze the data of all of the combined CHEM-571 data.
<br>
<br>
==Preparation of Adenosine Stock solution==
==Preparation of Adenosine Stock solution==
General Protocol
#In a clean 50mL volumetric flask, add 0.200g adenosine. Fill the volumetric flask with deionized H<sub>2</sub>O up until the white line.
#In a clean 50mL volumetric flask, add 0.200g adenosine. Fill the volumetric flask with deionized H<sub>2</sub>O up until the white line.
#*'''Note''': To ensure the full addition of all of the adenosine, a small amount of deionized H<sub>2</sub>O was placed on the weigh boat and a pipette was used to remove all of the H<sub>2</sub>O on the weigh boat.s
#*'''Note''': To ensure the full addition of all of the adenosine, a small amount of deionized H<sub>2</sub>O was placed on the weigh boat and a pipette was used to remove all of the H<sub>2</sub>O on the weigh boat.s
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#Pour the solution into a 50mL falcon tube for later storage.
#Pour the solution into a 50mL falcon tube for later storage.
#Dilute the solution by taking the necessary volume of the solution (using an automated pipette), placing the solution into the appropriately sized volumetric flask, and add deionized H<sub>2</sub>O.
#Dilute the solution by taking the necessary volume of the solution (using an automated pipette), placing the solution into the appropriately sized volumetric flask, and add deionized H<sub>2</sub>O.
<br>
*Needed Stock Solutions for Adenosine
<br>
{|style="width:700px"
|<u>Adenosine solution concentrations (M)</u>
|-
|3.00x10<sup>-5</sup>
|-
|2.50x10<sup>-5</sup>
|-
|2.00x10<sup>-5</sup>
|-
|1.50x10<sup>-5</sup>
|-
|1.00x10<sup>-5</sup>
|-
|0.50x10<sup>-5</sup>
|-
|RANDOM
|}
<br>
<br>
* Stock Solution Preparation for Adenosine   
* Stock Solution Preparation for Adenosine   
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  |}
  |}
<br>
<br>
==Preparation of Inosine Stock Solutions==
General Protocol
#In a clean 25mL volumetric flask, add 0.1347g inosine. Fill the volumetric flask with deionized H<sub>2</sub>O up until the white line.
#*'''Note''': To ensure the full addition of all of the inosine, a small amount of deionized H<sub>2</sub>O was placed on the weigh boat and a pipette was used to remove all of the H<sub>2</sub>O on the weigh boat.s
#Cap the volumetric flask and shake the volumetric flask vigorously until the inosine is completely dissolved.
#*'''Note''':The solution should be shaken vigorously for a minimum of 3 minutes.
#Pour the solution into a 50mL falcon tube for later storage.
#Dilute the solution by taking the necessary volume of the solution (using an automated pipette), placing the solution into the appropriately sized volumetric flask, and add deionized H<sub>2</sub>O.
<br>
*Needed Concentrations for Inosine Solutions
{|style="width:700px"
|<u>Inosine solution concentrations (M)</u>
|-
|4.80x10<sup>-5</sup>
|-
|4.00x10<sup>-5</sup>
|-
|3.20x10<sup>-5</sup>
|-
|2.40x10<sup>-5</sup>
|- 
|1.60x10<sup>-5</sup>
|-
|0.80x10<sup>-5</sup>
|-
|RANDOM
|}
<br>
<br>
* Stock Solution Preparation for Inosine 
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Total Mass [g]'''
| align="center" style="background:#f0f0f0;"|'''Stock Volume [mL]'''
| align="center" style="background:#f0f0f0;"|'''Concentration ['''M''']'''
|-
| 0.1347||25||0.02|||
|}
<br>
*Serial Dilution of Inosine Stock Solution
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Dilution'''
| align="center" style="background:#f0f0f0;"|''''''M<sub>1'''[mol/L]'''
| align="center" style="background:#f0f0f0;"|''''''V<sub>1''' [mL]'''
| align="center" style="background:#f0f0f0;"|''''''M<sub>2'''[mol/L]'''
| align="center" style="background:#f0f0f0;"|''''''V<sub>2''' [mL]'''
|-
| 1||0.02||0.24||4.8E-4||10||
|-
| 2||4.8E-4||1.0||4.8E-5||10||
|-
| 3||4.8E-4||0.833||4.0E-5||10||
|-
| 4||4.8E-4||0.667||3.2E-5||10||
|-
| 5||4.8E-4||0.5||2.4E-5||10||
|-
| 6||4.8E-4||0.333||1.6E-5||10||
|-
| 7||4.8E-4||0.1667||0.8E-5||10||
|-
| 8||1.5E-4||0.25||1.2E-5||10||
|}
<br>
==UV/vis==
'''Figure 1. Absorbance of Standard Adenosine Solutions'''
<br>
[[Image:Correctedabsadenosine09042013rerunzem.png]]
<br>
*Maximum absorbance occurred at 260nm for each adenosine concentration
'''Figure 2. Calibration Curve of Adenosine Solutions'''
<br>
[[Image:Calibrationcurveadenosinezem09082013.png]]
<br>
'''Figure 3. Absorbance of Standard Inosine Solutions'''
<br>
[[Image:Correctedabsinosinezem09042013.png]]
<br>
*The maximum absorbance of each solution occurred at 249nm.
'''Figure 4. Calibration Curve of Inosine Solutions'''
<br>
[[Image:Calibrationcurveinosinestdszem09082013.png]]
==Notes==
==Notes==
*Originally the adenosine stock solution was prepared  
*Originally the adenosine stock solution was prepared as 0.03M in 25mL. However, the adenosine was not soluble in such a small volume so the original adenosine solution was poured into a 50mL volumetric flask and rinsed thoroughly with deionized H<sub>2</sub>O.
*Adenosine samples of 1.0E-5M and 3.0E-5M were re-prepared and re-run because these points were outliers on our original calibration curve.
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
|}
|}


__NOTOC__
__NOTOC__

Revision as of 19:10, 8 September 2013

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Objectives

  1. Prepare adenosine solutions of various concentrations to determine the molar absorptivity of adenosine.
  2. Prepare inosine solutions of various concentrations to determine the molar absorptivity of inosine.
  3. Prepare calibration curves for adenosine and inosine.
  4. Analyze the data of all of the combined CHEM-571 data.


Preparation of Adenosine Stock solution

General Protocol

  1. In a clean 50mL volumetric flask, add 0.200g adenosine. Fill the volumetric flask with deionized H2O up until the white line.
    • Note: To ensure the full addition of all of the adenosine, a small amount of deionized H2O was placed on the weigh boat and a pipette was used to remove all of the H2O on the weigh boat.s
  2. Cap the volumetric flask and shake the volumetric flask vigorously until the adenosine is completely dissolved.
    • Note:The solution should be shaken vigorously for a minimum of 5 minutes.
  3. Pour the solution into a 50mL falcon tube for later storage.
  4. Dilute the solution by taking the necessary volume of the solution (using an automated pipette), placing the solution into the appropriately sized volumetric flask, and add deionized H2O.


  • Needed Stock Solutions for Adenosine


Adenosine solution concentrations (M)
3.00x10-5
2.50x10-5
2.00x10-5
1.50x10-5
1.00x10-5
0.50x10-5
RANDOM


  • Stock Solution Preparation for Adenosine
Total Mass [g] Stock Volume [mL] Concentration [M]
0.2006 50 0.015


  • Serial Dilution of Adenosine Stock Solution
Dilution 'M1'[mol/L] 'V1' [mL] 'M2'[mol/L] 'V2' [mL]
1 0.015 0.1 1.5E-4 10
2 0.015 0.25 3.0E-5 10
3 1.5E-4 1.667 2.5E-5 10
4 1.5E-4 1.333 2.0E-5 10
5 1.5E-4 1.0 1.5E-5 10
6 1.5E-4 0.667 1.0E-5 10
7 1.5E-4 0.333 0.5E-5 10
8 1.5E-4 0.5 0.75E-5 10


Preparation of Inosine Stock Solutions

General Protocol

  1. In a clean 25mL volumetric flask, add 0.1347g inosine. Fill the volumetric flask with deionized H2O up until the white line.
    • Note: To ensure the full addition of all of the inosine, a small amount of deionized H2O was placed on the weigh boat and a pipette was used to remove all of the H2O on the weigh boat.s
  2. Cap the volumetric flask and shake the volumetric flask vigorously until the inosine is completely dissolved.
    • Note:The solution should be shaken vigorously for a minimum of 3 minutes.
  3. Pour the solution into a 50mL falcon tube for later storage.
  4. Dilute the solution by taking the necessary volume of the solution (using an automated pipette), placing the solution into the appropriately sized volumetric flask, and add deionized H2O.


  • Needed Concentrations for Inosine Solutions
Inosine solution concentrations (M)
4.80x10-5
4.00x10-5
3.20x10-5
2.40x10-5
1.60x10-5
0.80x10-5
RANDOM



  • Stock Solution Preparation for Inosine
Total Mass [g] Stock Volume [mL] Concentration [M]
0.1347 25 0.02


  • Serial Dilution of Inosine Stock Solution
Dilution 'M1'[mol/L] 'V1' [mL] 'M2'[mol/L] 'V2' [mL]
1 0.02 0.24 4.8E-4 10
2 4.8E-4 1.0 4.8E-5 10
3 4.8E-4 0.833 4.0E-5 10
4 4.8E-4 0.667 3.2E-5 10
5 4.8E-4 0.5 2.4E-5 10
6 4.8E-4 0.333 1.6E-5 10
7 4.8E-4 0.1667 0.8E-5 10
8 1.5E-4 0.25 1.2E-5 10


UV/vis

Figure 1. Absorbance of Standard Adenosine Solutions

  • Maximum absorbance occurred at 260nm for each adenosine concentration

Figure 2. Calibration Curve of Adenosine Solutions

Figure 3. Absorbance of Standard Inosine Solutions

  • The maximum absorbance of each solution occurred at 249nm.

Figure 4. Calibration Curve of Inosine Solutions

Notes

  • Originally the adenosine stock solution was prepared as 0.03M in 25mL. However, the adenosine was not soluble in such a small volume so the original adenosine solution was poured into a 50mL volumetric flask and rinsed thoroughly with deionized H2O.
  • Adenosine samples of 1.0E-5M and 3.0E-5M were re-prepared and re-run because these points were outliers on our original calibration curve.


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