Waters Advanced Purification Glass Columns - Manual (Page 3)

[ Care and Use ManUal ]
Waters Advanced Purification Columns 3
Using the coarse/fine adjustment slide
The AP column features a coarse/fine adjustment slide to make
plunger adjustment fast and easy. Make sure the slide is in the down
position (against the fine adjustment knob) before pressurizing the
column, Figure 3 shows:
1. Deactivating the inlet O-ring seal
2. Using the slide for coarse adjustment
3. Using the slide for fine adjustment and locking
The slide must engage the locking hex during operation to
prevent the inlet plunger from unscrewing under pressure.
4. Activating the O-ring seal
Figure 3: Adjusting the Plunger Downward
c. Packing the Column
The AP Column is shipped unpacked. The Waters AP Column is
compatible with all commonly used silica- and resin-based packing
materials. Repack the column when performance deteriorates or
when a new method requires a different packing material.
When you repack an AP Column, empty the column first (see
“Emptying the Column”), then proceed to the correct section for
slurry-packing or dry-packing the column.
Column volumes
When you pack the column, you must calculate the column volume
from the length of the packed bed you will use in your method.
Column volumes are:
Short beds
To pack a column with a shorter bed, replace the outlet connector
with a second inlet connector (see Section V, “Ordering
Information”). Inlet connectors have a 40 mm range of movement,
allowing you to reduce column volume to as little as 3 mL. The
minimum bed length for a 100 mm long column with two inlet
connectors is 20 mm.
Slurry-packing the column
Amount of packing material required
To determine the amount of packing material required, multiply the
column volume, V (mL), by the bulk density of the packing material,
d (g/ml):
Amount of packing material required = V x d
Determining density
To determine the density of the packing material:
1. Slurry 1 g of packing material in 10 mL of buffer and allow the mate-
rial to settle.
2. Note the volume of the settled material
3. Calculate density by dividing the weight of the material by the settled
volume.
Column length Column volume
100 mm 8 to 8 mL
200 mm 13 to 16 mL
300 mm 21 to 24 mL
600 mm 45 to 48 mL
Packing Material Bulk Density of the Packing Material
Waters Protein-Pak™ HR 1.2 g/mL
Waters AccellPlus™ silica-based
packing materials
0.5 g/mL
Other packing materials Use procedure below to determine density