YMC Europe GmbH
Operation
YMC Pilot
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Low pressure chromatography column
Check that no air bubbles are below the frit.
16. Turn the cylinder screws with the hex-wrench through the closing disc into the
column disc. This way, the column is closed.
17. Tighten the O-ring of the adjustable end piece.
18. Open the valve in the tubing on the adjustable end piece.
19. Turn the piston of the adjustable end piece in the down direction until the
tubing is completely filled with liquid.
The tubing is now free of air.
20. Connect the pump with the tubing of the adjustable end piece.
21. Open the valve in the tubing of the fixed end piece.
22. Start the flow (top-down).
23. Increase the flow rate stepwise until the desired pressure / flow rate is
reached. Wait after every step until the pressure is constant. Afterwards, the
flow rate can be further increased.
Consider the manufacturer´s recommendations for the packing material.
The flow rate should be considerably (>20%) higher than the required
operating flow rate.
The column bed is visually compressed.
24. Check and mark the bed length frequently.
25. If the final bed length is reached, stop the pump.
26. Turn the piston of the adjustable end piece in the down direction until the
column bed is reached.
27. Start the pump.
28. Check, if the column bed is further compressed. A dead volume would occur.
29. If a dead volume occurs, stop the pump and repeat step 23 to 26.
30. If no dead volume occurs, the pilot column is released Chapter 6.5, p. 32.
31. Equilibrate the glass column with the desired eluent and test the column
performance.
6.5
Column test and release criteria
The column test and the release criteria are selected with regards to the later
application requirements. The preparation of the required specifications for the final
use of a column or for the purification of a target molecule is determined individually.
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