Objective
- Prepare a new batch of fibers.
- Prepare HPLC Mobile Phase.
- Learn how to use HPLC.
- Create UV-Vis calibration curve for Acetylsalicylic acid
Protocol
- Fiber samples were prepared on a 5x scale according to the protocol described on February 16th.
- A 25:75 Acetonitrile:water solution was made with a pH adjusted to 2.6 by Phosphorous acid.
- Similarly, a 75:25 Acetonitrile:water solution was made with a pH adjusted to 2.6 by Phosphorous acid.
- Both solutions were filtered through a .2μm filter to remove any particulates before use in HPLC.
- Matt and I created a UV-Vis calibration Curve for Acetylsalicylic acid using 505 μM, 252.5 μM, 126.25 μM, and 63 μM standards. The absorbance was analyzed at 297 nm.
- Dr. Fox and I equilibrated the HPLC with the new solvents, then we all ran an HPLC separation on the 1.01mM stock solution of acetylsalicylic acid as a means to learn how the instrument is operated.
Calibration Curve
The graph above relates the concentration of acetylsalicylic acid in an aqueous solution to it's absorbance in UV-Vis.
Notes
- It should be noted that HPLC grade solvents were not used to make the mobile phase solutions.
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