- Setup AgNP and AuNP synthesis reactions for SPI, WPI, Casein, Pea Protein and Brown Rice Protein
Materials
- Used same nutritional proteins on 02/02/16
- Protein and gold stocks prepared fresh every day
- Ocean Optics UV Vis was used to record UVVIS Spectra of Casein AgNPs. The other machine broke and is unusable.
- Integration time= 10ms and Average =100.
- A water blank in the same quartz cuvette was subtracted from the spectra before running any samples.
Procedure
- Reducing stock protein concentrations (and thus concentrations in reaction mix) to increase metal/protein ratio to spur NP formation as we saw protein aggregate and nanofiber formation in previous reactions
- using citrate instead of fructose as reducing agent
- comparing citrate to no citrate samples, with citrate as only variable
Detailed Reaction Setup shown Below (4 hours 80˚C, 1ml rxns)
| '
|
Tube #
|
2.5mM HAuCl4 (µL)
|
4.375mMAgNO3 (µL)
|
Protein (µL)
|
10 mM Citrate (µL)
|
Water (µL)
|
Au/Ag:Protein
|
Notes
|
|
1 |
40 |
0 |
560 |
0 |
400 |
|
no change
|
| .5mg/mL Casein |
2 |
40 |
0 |
560 |
20 |
380 |
|
|
| grams weighed: 5.06mg |
3 |
40 |
0 |
200 |
0 |
760 |
|
|
| Stock: 10mL |
4 |
40 |
0 |
200 |
20 |
740 |
|
|
| -reduce from 2mg/ml to .5mg/ml returning previous working concentration |
5 |
0 |
40 |
560 |
0 |
400 |
|
all yellow soln, no particles
|
|
6 |
0 |
40 |
560 |
0 |
400 |
|
UV VIS
|
|
7 |
0 |
40 |
560 |
0 |
400 |
|
|
|
8 |
40 |
0 |
560 |
0 |
400 |
|
pink solutions w/ purple fibers on walls
|
|
9 |
40 |
0 |
560 |
20 |
380 |
|
pink soln with purple aggreagte on bottom
|
| 0.25mg/mL SPI |
10 |
40 |
0 |
360 |
0 |
600 |
|
pink solution w/ purple fibers on walls
|
| grams weighed: xmg |
11 |
40 |
0 |
360 |
20 |
580 |
|
pink soln with purple aggreagte on bottom
|
| Stock: xmL |
12 |
40 |
0 |
200 |
0 |
760 |
|
pink soln with purple aggreagte on bottom
|
|
13 |
40 |
0 |
200 |
20 |
740 |
|
UV VIS pink soln with purple aggreagte on bottom
|
| -change from 2/9/16, reduced concentration from .75mg/ml to .25mg/ml |
14 |
0 |
40 |
560 |
0 |
400 |
|
No change 14-19
|
|
15 |
0 |
40 |
560 |
20 |
380 |
|
|
|
16 |
0 |
40 |
360 |
0 |
600 |
|
|
|
17 |
0 |
40 |
360 |
20 |
580 |
|
|
|
18 |
0 |
40 |
200 |
0 |
760 |
|
|
|
19 |
0 |
40 |
200 |
20 |
740 |
|
|
|
20 |
40 |
0 |
560 |
0 |
400 |
|
20-25 all pink solutions
|
|
21 |
40 |
0 |
560 |
20 |
380 |
|
some purple aggregate on walls
|
|
22 |
40 |
0 |
360 |
0 |
600 |
|
|
| .25mg/mL Whey Protein |
23 |
40 |
0 |
360 |
20 |
580 |
|
|
| grams weighed: 10mg |
24 |
40 |
0 |
200 |
0 |
760 |
|
purple aggregate at bottom of tube
|
| Stock: 10mL |
25 |
40 |
0 |
200 |
20 |
740 |
|
UV VIS
|
|
26 |
0 |
40 |
560 |
0 |
400 |
|
No change 26-31
|
| -change from 2/9/16, reduced concentration from .75mg/ml to .25mg/ml |
27 |
0 |
40 |
560 |
20 |
380 |
|
|
|
28 |
0 |
40 |
360 |
0 |
600 |
|
|
|
29 |
0 |
40 |
360 |
20 |
580 |
|
|
|
30 |
0 |
40 |
200 |
0 |
760 |
|
|
|
31 |
0 |
40 |
200 |
20 |
740 |
|
|
|
32 |
40 |
0 |
560 |
0 |
400 |
|
No change 32-37
|
|
33 |
40 |
0 |
560 |
20 |
380 |
|
|
| 0.25mg/mL Pea Protein |
34 |
40 |
0 |
360 |
0 |
600 |
|
|
| grams weighed: 10.38mg |
35 |
40 |
0 |
360 |
20 |
580 |
|
|
| Stock: 10.3mL |
36 |
40 |
0 |
200 |
0 |
760 |
|
|
|
37 |
40 |
0 |
200 |
20 |
740 |
|
|
| -dissolve in acid instead of base |
38 |
0 |
40 |
560 |
0 |
400 |
|
38-43 all yellow soln, silver nanoparticles
|
|
39 |
0 |
40 |
560 |
20 |
380 |
|
UVVIS on All
|
|
40 |
0 |
40 |
360 |
0 |
600 |
|
|
|
41 |
0 |
40 |
360 |
20 |
580 |
|
|
|
42 |
0 |
40 |
200 |
0 |
760 |
|
|
|
43 |
0 |
40 |
200 |
20 |
740 |
|
|
|
44 |
40 |
0 |
560 |
0 |
400 |
|
no change
|
|
45 |
40 |
0 |
560 |
20 |
380 |
|
purple AUNPs
|
| 0.25mg/mL Rice Protein |
46 |
40 |
0 |
360 |
0 |
600 |
|
no change
|
| grams weighed: 10mg |
47 |
40 |
0 |
360 |
20 |
580 |
|
purple AUNPs
|
| Stock: 10mL |
48 |
40 |
0 |
200 |
0 |
760 |
|
no change
|
|
49 |
40 |
0 |
200 |
20 |
740 |
|
UVVIS Purple AUNPs
|
| -this needs to be dissolved by base, particles float to the bottom |
50 |
0 |
40 |
560 |
0 |
400 |
|
no change
|
| -Did not dissolve. We didn't add* try next week |
51 |
0 |
40 |
560 |
20 |
380 |
|
slightly yellow
|
|
52 |
0 |
40 |
360 |
0 |
600 |
|
no change
|
|
53 |
0 |
40 |
360 |
20 |
580 |
|
slightly yellow
|
|
54 |
0 |
40 |
200 |
0 |
760 |
|
no change
|
|
55 |
0 |
40 |
200 |
20 |
740 |
|
slightly yellow
|
Results
Figure 1: UVVis Spectra for Pea Protein AgNP Samples With and Without Citrate (synthesized on 02/10/2016)
800px
Figure 2: UVVis Spectra for Casein AgNP Samples in Triplicate (synthesized on 02/10/2016)
Figure 3: UVVis Spectra for WPI AuNP Samples by Protein Concentration (synthesized on 02/010/2016)
-Only 2 of the WPI-Au samples with citrate formed nanoparticles, the WPI-Au samples without citrate formed nanofibers which clung to walls.
Figure 4: AuNP UV-Vis Spectra for our Red AuNPs (synthesized on 02/10/2016)
-This shows UVVIS Spectra for SPI-Au sample with lowest protein concentration tested and highest concentration of citrate. All other SPI-Au samples formed protein aggregate, fibers or both.
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