User:Benjamin Friedel/Notebook/CHEM 471/2015/10/07
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Ocean Optics Spectrophotometry was used to measure absorbance of an AuNP fibers sample over time as it was incubated with the protease, alpha-chymotrypsin. This was performed to track how alpha-chymotrypsin digests lysozyme based AuNP fibers.
AuNP Fiber Sample Preparation
Incubation Mix Preparation
In this experiment, the blank or control data set is considered to be the first data point collected after the initial data point collection. To correct for this blank data (without protease), the absorbance values at each wavelength were subtracted from the absorbance data for each 2 minute measurement after addition of protease (alpha chymotrypsin). Then, to correct for instrumental noise, the absorbance values for the last 60nm for each 2min scan were averaged individually and subtracted from that individual scan.
Figure 1 above shows the absorbance values for each scan after the first 2min at 530nm are shown as time progressed. This is because AuNP fibers were previously determined to have an absorbance max at 530nm, so their breakdown into solution upon digestion of AuNP fibers by alpha-chymotrypsin was monitored. This figure shows that before the protease was added, the fibers have a stronger absorbance. After protease addition the quick decrease in absorbance that reached a lower limit around 0.8 suggests that the fibers are degraded into constituents that do not absorb at 530nm and that this occurs relatively quickly and comprehensively.