User:Moira M. Esson/Notebook/CHEM-571/2013/09/11: Difference between revisions

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* Determine number of atoms of gold in the BSA and citrate gold solutions using the prepared gold calibration curve and the diameter size of the gold particles in the BSA and citrate solutions(this can be determined from the UV/vis samples run of these on [[User:Moira M. Esson/Notebook/CHEM-571/2013/08/28|08/28/2013]].
* Determine number of atoms of gold in the BSA and citrate gold solutions using the prepared gold calibration curve and the diameter size of the gold particles in the BSA and citrate solutions(this can be determined from the UV/vis samples run of these on [[User:Moira M. Esson/Notebook/CHEM-571/2013/08/28|08/28/2013]].
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==Adenosine Deaminase Bradford assay==
==Adenosine Deaminase Bradford assay==
*All standard solutions were re-prepared using the procedure described on [[User:Moira M. Esson/Notebook/CHEM-571/2013/09/10|09/10/2013]].
*All standard solutions were re-prepared using the procedure described on [[User:Moira M. Esson/Notebook/CHEM-571/2013/09/10|09/10/2013]].
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'''Figure 2. Calibration Curve for ADA standard solutions at 596nm'''
'''Figure 2. Calibration Curve for ADA standard solutions at 596nm'''
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[[Image:Calibrationcurvebradfordassay09112013zem.png]]
[[Image:Calibrationcurvebradfordassay09112013zem.png]]
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==Atomic Absorption==
==Atomic Absorption==
'''Figure 3. Calibration curve of gold nanoparticles'''
'''Figure 3. Calibration curve of gold nanoparticles'''
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[[Image:AAcalibrationcurvechem57109112013zem.png]]
[[Image:AAcalibrationcurvechem57109112013zem.png]]
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'''Figure 4. CHEM-571 Group calibration curve of gold nanoparticles'''
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[[Image:GroupcalibrationcurveAAgoldchem571zem.png]]
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'''Table 1. Grubb's test for prepared group calibration curve of gold standards'''
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{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Concentration(μg/mL)'''
| align="center" style="background:#f0f0f0;"|'''Suspected Outliers(Abs)'''
| align="center" style="background:#f0f0f0;"|'''G<sub>exp</sub>'''
|-
| 5||0.1164||1.288081628
|-
| 10||0.2102||1.140265886
|-
| 15||0.2856||1.216520053
|-
| 20||0.4572||1.571958215
|-
| 25||0.5377||1.029873242
|-
| 25||0.4619||1.014767716
|}
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*With N=4, G<sub>critical</sub> at 95% is 1.46. The absorbance of 0.4572 for 20μg/mL can be discarded with 95% certainty.
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'''Table 2. Concentration and atom number of gold in prepared solutions'''
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{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''Concentration gold(ug/mL)'''
| align="center" style="background:#f0f0f0;"|'''Atoms gold'''
|-
| BSA||0.015551764||4.77E+13
|-
| Citrate-AuNP 1/10 dilution||0.006918789||2.11529E+13
|}




Revision as of 10:32, 18 September 2013

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Objectives

  • Run Au-nanoparticles samples prepared 09/10/2013 on the AA.
  • Run BSA solution prepared on 08/28/2013 on the AA.
  • Prepare a 1/10 dilution of the Au-citrate solution prepared 08/28/2013 and run this sample on the AA.
  • Prepare calibration curve for Au-nanoparticle solutions and determine the concentration of Au in the BSA and citrate solutions.
  • Re-run bradford assay of adenosine deaminase that was unsuccessful on 09/10/2013.
  • Determine number of atoms of gold in the BSA and citrate gold solutions using the prepared gold calibration curve and the diameter size of the gold particles in the BSA and citrate solutions(this can be determined from the UV/vis samples run of these on 08/28/2013.


Adenosine Deaminase Bradford assay

  • All standard solutions were re-prepared using the procedure described on 09/10/2013.


Figure 1. Absorption spectra of ADA standard solutions prepared by Bradford Assay Trial #4

  • This trial has the correct shape. It was determined that the machine was not subtracting the blank. The blank was maunally subtracted to produce the following spectrum. Maximum absorption was observed at 596nm.
  • The solutions prepared today seemed slightly more blue than the solutions prepared on 09/10/2013. Another potential issue may have been the addition of the Bio-Rad.


Figure 2. Calibration Curve for ADA standard solutions at 596nm

Atomic Absorption

Figure 3. Calibration curve of gold nanoparticles

Figure 4. CHEM-571 Group calibration curve of gold nanoparticles

Table 1. Grubb's test for prepared group calibration curve of gold standards

Concentration(μg/mL) Suspected Outliers(Abs) Gexp
5 0.1164 1.288081628
10 0.2102 1.140265886
15 0.2856 1.216520053
20 0.4572 1.571958215
25 0.5377 1.029873242
25 0.4619 1.014767716


  • With N=4, Gcritical at 95% is 1.46. The absorbance of 0.4572 for 20μg/mL can be discarded with 95% certainty.


Table 2. Concentration and atom number of gold in prepared solutions

' Concentration gold(ug/mL) Atoms gold
BSA 0.015551764 4.77E+13
Citrate-AuNP 1/10 dilution 0.006918789 2.11529E+13




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