Objective
- Create and analyze 2 hour incubation samples
- None of the BSA samples produced fibers, so we are no longer using
Protocol
- At 10:30 this morning, I came in and added Rhodamine B to nine of the lysozyme fiber samples created yesterday. Three samples with concentrations of 2, 5, and 10 μM were created. For the volumes of stock Rhodamine B used, see Nicole's notebook.
- These samples were analyzed by centrifuging the tubes for 2 minutes to get the fibers to move to the bottom of the test tube so that supernatant could be analyzed.
- 3 mL samples were then analyzed using UV-Vis. Spectra were created from 300-800 nm using polystyrene cuvettes.
- A standard curve was created by placing Rhodamine B in fiber solutions then immediately removing supernatant once thoroughly mixed through. Standard concentrations used were 1, 2, 4, 5, 8, and 10 μM.
Results
Figure 1: The graph above is a UV-Vis calibration curve relating absorbance of Rhodamine B at 556 nm when dissolved in AuNP fiber supernatant to the concentration of Rhodamine B. The equation of the line is included on the graph. The high R2 value indicates the strong fit of the trendline/equation to the data.
Figure 2: The table above shows the raw and calculated data including the peak absorbance, the concentration that this absorbance corresponds to, the change in concentration after incubation, and the percent change in concentration.
Figure 3: The chart above displays the percent change in concentration of Rhodamine B in solution after incubation for 2 hours. Blue bars correspond to 10 μM initial Rhodamine concentrations, red bars correspond to 5 μM initial Rhodamine concentrations, and green bars correspond to 2 μM initial Rhodamine concentrations. As expected, the concentration of Rhodamine B in solution decreases indicating that Rhodamine B is likely being incorporated into the AuNP fibers.
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