User:Jacob Esenther/Notebook/Chem 571/2014/09/30

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Created a create a Ca2+ calibration curve and began a new dialysis


  1. CaCl2 Calibration Curve
    • Prepare 2 mL 50 mM CaCl2 (if you read ahead, you can use your CaCl2 calibration stock)
    • Prepare 2 mL 500 μM CaCl2 (if you read ahead, you can use a CaCl2 calibration dilution)
    • Prepare 2 mL 50 mM NaCl (if you read ahead, you can use your NaCl calibration stock)
  2. Cl- calibration curve
    • Prepare 25 mL 50 mM NaCl using HPLC water
      • We calculated that 0.07305g NaCl was necessary
    • Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM NaCl
    • Measure Cl- using ISE
    • Try measuring 50 μM and 50 mM of other salts to check for interferences & ion-pairing
    • Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution.
  3. Dialysis
    • Prepare 10 mL 0.5 g/L Lysozyme
    • Cut a piece of 3500 MWCO flat dialysis tubing, wet it, and insert it in the 5-well dialysis chamber
    • Clamp the dialysis chamber shut
    • Fill one side of each well with 1 mL of your 0.5 g/L Lysozyme solution
      • Transfer 1 mL to a plastic test tube using a pippetter, then
      • Carefully fill each well with with a glass Pasteur pipette
      • You may need to tip the chamber on an angle to reduce water surface tension
    • Fill the other side of the well with one of the following solutions
      • HPLC water, 0.25 mM HCl, 50 mM CaCl2, 500 μM CaCl2, or 50 mM NaCl
      • Each well has a different solution, which will be tested tomorrow
    • Insert top screws to prevent leaking/evaporation


  • For the 0.5 g/L Lysozyme, 0.00570g Lysozyme used.
  • For NaCl serial dilution 0.0713g NaCl was placed in 25 mL of water
NaCl Calibration Curve Conc. (mM) mV
50 -24
5 28.5
0.5 88.2
0.05 128.8
0.005 152.8

Polyvinyl Alcohol Films with Absorbed Malachite Green Concentrations; 0.2, 0.5, 1.1, 1.4, 1.7, and 2.0 ppm


  • For the 0.25 mM HCl, a new solution was made containing HPLC water rather than DI water