User:Benjamin Friedel/Notebook/472 CHEM/2016/01/26

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Objective

Todays objective is to prepare gold and silver nanoparticles using pure casein and soy protein isolate as capping and reducing agents

Materials

Stock Solutions

  1. SPI (ARCON S 066400)
  1. Casein (amresco E666-500G)
    1. note store in refrigerator
  1. Fructose
  1. Choloroauric Acid (REM Chemicals)
    1. note store in desiccator, sensitive to metals
  1. Silver Nitrate (CMI 7985)
    1. note store in desiccator, sensitive to light

Procedure

  • Gold Nanoparticles

Procedure to make the gold nanoparticles was based off of Dr. Hartings' notebook. Instead of lysozyme, Casein (an estimated MW at 22kD) was used. Then the concentration of Casein for 20uM was converted to g/ml, and this concentration was applied to the soy protein isolate (SPI) which does not have a clear MW.

The reaction spreadsheet is shown below. After creating the reaction mixes, samples were heated at 80˚C for 4 hours in the HERATHERM oven in Dr. Harting's Lab in Beeghly (on AU campus). This reaction cycle also consisted of 30 minutes of heating and cooling beforehand. Samples were retrieved the next day (1/27/16), checked for color change and then covered in tin foil completely (to prevent oxidation by light of potentially unreacted silver nitrate).


1/27/2016 ' ' ' ' ' ' ' ' '
1mL total reaction 2.5mM HAuCl4 2.5mMAgNO3 .44mg/ml SPI .44mg/ml Casein 2.5mM Fructose Water Au/Ag:Protein Notes
1 40µL 0 0 560µL 0 400µL 1:15 no color change
2 40µL 0 0 440µL 0 520µL 1:11
3 40µL 0 0 360µL 0 600µL 1:09
4 40µL 0 0 280µL 0 680µL 1:07
5 40µL 0 0 200µL 0 760µL 1:05
6 0 40µL 0 560µL 0uL 400µL 1:15 all gold/bright yellow except for #14 which was lighter
7 0 40µL 0 560µL 20uL 380µL 1:15
8 0 40µL 0 560µL 40uL 360µL 1:15
9 0 40µL 0 360uL 0uL 600µL 1:09
10 0 40µL 0 360µL 20uL 580µL 1:09
11 0 40µL 0 360µL 40uL 560µL 1:09
12 0 40µL 0 200uL 0uL 760µL 1:05
13 0 40µL 0 200µL 20uL 740µL 1:05
14 0 40µL 0 200µL 40uL 720µL 1:05
15 40µL 0 560µL 0 0 400µL 1:15 Made fibers so need higher concentration of protein
16 40µL 0 440µL 0 0 520µL 1:11 purple fibers at bottom
17 40µL 0 360µL 0 0 600µL 1:09
18 40µL 0 280µL 0 0 680µL 1:07
19 40µL 0 200µL 0 0 760µL 1:05
20 0 40µL 560µL 0 0uL 400µL 1:15 No color change
21 0 40µL 560uL 0 20uL 380µL 1:15
22 0 40µL 560µL 0 40uL 360µL 1:15
23 0 40µL 360uL 0 0uL 600µL 1:09
24 0 40µL 360µL 0 20uL 580µL 1:09
25 0 40µL 360µL 0 40uL 560µL 1:09
26 0 40µL 200uL 0 0uL 760µL 1:05
27 0 40µL 200µL 0 20uL 740µL 1:05
28 0 40µL 200µL 0 40uL 720µL 1:05
Controls 29 40µL 0 0 0 0 960µL No color change
30 0 40µL 0 0 0 960µL
-oven started 4:35pm 1/27, removed and covered 10:30am 1/28


  • Silver nanoparticles

Results

  1. Silver nanoparticles with casein were created as seen by a color change from clear to yellow, as well as a clear and solitary peak at 420nm in UV Vis.