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

Biomaterials Design Lab

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Objective Created a create a Ca2+ calibration curve and began a new dialysis Description 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) 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. 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 Data 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 Notes For the 0.25 mM HCl, a new solution was made containing HPLC water rather than DI water