User:Khyra A. Neal/Notebook/Chem 571/2014/09/30

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September 30, 2014

Tasklist

1. Calibration Curve for CaCl2

  • Prepare 25 mL 50 mM CaCl2 using HPLC water
    • 0.13875 g CaCl2 in 25 mL HPLC water
  • Prepare 25 mL of each using serial dilutions: 5 mM, 500 μM, 50 μM, and 5 μM CaCl2 from 50 mM stock CaCl2
    • Dilute all solutions to 25 mL with HPLC water.
  • Measure Ca2+ using ISE
CaCl2 concentration Potential (mV)
5μM -44.6
50μM 8.2
500μM 39.5
5mM 63.5
50mM 86.6

Image:CaCl2.jpg

  • Measure HPLC water, DI water, tap water, and your lysozyme stock solution.
Substance Potential (mV)
Tap water 51.3
HPLC water -23.0
DI water -51.5
Lysozyme 0.5g/L -19.1
  • Measure 50 mM and 50 mM of other salts to check for interferences
CaCl2 concentration Cu(II)Cl2 concentration KCl concentration Potential (mV)
50mM 050mM86.4
50mM 50mM080.5


2. Calibration Curve for NaCl

  • Prepare 25 mL of 50 mM NaCl using HPLC water
    • 0.07305 g NaCl in 25 mL HPLC water
  • Prepare 25 mL of each using serial dilutions: 5 mM, 500 μM, 50 μM, and 5 μM NaCl from 50 mM NaCl stock solution
    • Dilute all solutions to 25 mL with HPLC water
  • Measure Cl- using ISE
NaCl concentration Potential (mV)
5μM 146.5
50μM 129.6
500μM 81.3
5mM 27.5
50mM -23.9

Image:NaCl.jpg

  • Measure HPLC water, DI water, tap water, and your lysozyme stock solution.
Substance Potential (mV)
Tap water 65.5
HPLC water 163.5
DI water 158.8
Lysozyme 0.5g/L 89.4


3. Try measuring 50 mM and 50 mM of other salts to check for interferences

NaCl concentration NaSO4 Na2HPO42- Potential (mV)
50mM 50 mM0-27.0
50mM 050mM-30.8


Dialysis

  • Prepare 10 mL 0.5 g/L Lysozyme
    • 0.005 g Lysozyme in 25 mL HPLC water using volumetric flask
  • Prepare 0.25 mM HCl from 2.5 mM HCl stock solution
    • 10x dilution from stock
  • Cut a piece of 3500 MWCO flat dialysis tubing, wet it, and insert it in the 5-well dialysis chamber and clamp the dialysis chamber shut.
  • Fill one side of each well with 1 mL of 0.5 g/L Lysozyme solution
    • Measure out 1 mL with transfer pipette then add 1mL using Pasteur pipette to the wells
  • Fill the other side of the wells with the following solutions:
    • HPLC water, 0.25 mM HCl, 50 mM CaCl2, 500 μM CaCl2, and 50 mM NaCl
  • Insert screws to prevent leaving/evaporation
  • Place on low speed shaker overnight.



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