User:Michael S. Bible/Notebook/CHEM-671/690 Lab Notebook/2016/02/17

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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

UV-Vis calibration Curve Acetylsalicylic Acid.png

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.