1. Calculate heavy and light solutions to be made.
?methanol = 0.791
?ethylene glycol = 1.109
For PBD, PIB we need 0.86 to 0.92, go outside this range because column is not linear at ends, use 0.82 to 0.98.
0.82 is 90% methanol (1080 mL M, 120 mL E)
0.98 is 40% methanol (480 mL M, 720 mL E)
Prepare 1.2 L of each solution. And stir for a few hours.
2. The water level should be at the top of the black strips (pump inlet is at the top of the coils, need to cover).
3. Plug in the pump.
4. Turn on cooling water.
5. Wait until the temperature reaches 23oC exactly.
6. Put together valve that will connect both flasks and tubing that connects to the capillary column. Use a clamp to connect the joints without grease and place flasks on top of ledge.
7. Clean tube with water and hang upside down to dry.
8. Place black top on column.
9. Place column in water tank and secure with rubber bands.
10. Slide capillary tube through hole in column cap.
11. Grease the connection to the capillary column.
12. Fill the flasks (the light solution is closest to the capillary tube). The heavy solution should be either equal to or greater than the height of the light solution.
13. Start stirring the light solution (not too fast).
14. Position the capillary so it is not touching the bottom of the column and centered.
15. Open the valve between the flasks and equilibrate.
16. Open and close the valve to capillary tube to get the air out.
17. Open up the valve to the capillary tube, make sure the rate is not too fast.
18. Center the capillary tube if necessary.
19. Ideally about 15 minutes per step is a good rate, this can be changed in the beginning but not after.
20. Close off when column is full (will be well above top line).
21. Detach connection to capillary column.
22. Slowly remove capillary tube, keeping it straight.
23. Use foil scoop to remove beads from jars (do not scratch them) and drop in column.
24. Put cap without hole on column.
25. To add polymer, place a small amount on a slide and heat overnight. Freeze in liquid nitrogen and cut into small pieces. Add them into the column while frozen and let equilibrate.